potential impact of rapid blood culture testing for gram-positive … · 2019. 7. 30. · rdna or...

Research ArticlePotential Impact of Rapid Blood Culture Testing forGram-Positive Bacteremia in Japan with the VerigeneGram-Positive Blood Culture Test

Ken Kikuchi123 Mari Matsuda3 Shigekazu Iguchi13 Tomonori Mizutani3 KeiichiHiramatsu3 Michiru Tega-Ishii2 Kaori Sansaka2 Kenta Negishi2 Kimie Shimada2 JunUmemura2 Shigeyuki Notake4 Hideji Yanagisawa4 Hiroshi Takahashi5 Reiko Yabusaki6

Hideki Araoka6 and Akiko Yoneyama6

1Department of Infectious Diseases Tokyo Womenrsquos Medical University 8-1 Kawada-cho Shinjuku-ku Tokyo 162-8666 Japan2Sakakibara Heart Institute 1-16-1 Asahimachi Fuchu Tokyo 183-0003 Japan3Department of Infection Control Science Faculty of Medicine Juntendo University 2-1-1 Hongo Bunkyo-ku Tokyo 113-8421 Japan4Miroku Medical Laboratory Inc 659-2 Innai Saku Nagano 384-2201 Japan5East-West DiagnosticsTheranostics San Francisco CA 94105 USA6Department of Infectious Diseases Toranomon Hospital 2-2-2 Toranomon Minato-ku Tokyo 105-8470 Japan

Correspondence should be addressed to Ken Kikuchi kikuchikentwmuacjp

Received 18 October 2016 Revised 27 November 2016 Accepted 18 December 2016 Published 20 February 2017

Academic Editor Jose Ramon Blanco

Copyright copy 2017 Ken Kikuchi et al This is an open access article distributed under the Creative Commons Attribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Background Early detection of Gram-positive bacteremia and timely appropriate antimicrobial therapy are required for decreasingpatient mortality The purpose of our study was to evaluate the performance of the Verigene Gram-positive blood culture assay(BC-GP) in two special healthcare settings and determine the potential impact of rapid blood culture testing for Gram-positivebacteremia within the Japanese healthcare delivery system Furthermore the study included simulated blood cultures whichincluded a library of well-characterized methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci(VRE) isolates reflecting different geographical regions in JapanMethodsA total 347 BC-GP assays were performed on clinical andsimulated blood cultures BC-GP results were compared to results obtained by reference methods for genusspecies identificationand detection of resistance genes using molecular and MALDI-TOF MS methodologies Results For identification and detectionof resistance genes at two clinical sites and simulated blood cultures overall concordance of BC-GP with reference methods was327347 (94) The time for identification and antimicrobial resistance detection by BC-GP was significantly shorter compared toroutine testing especially at the cardiology hospital which does not offer clinical microbiology services on weekends and holidaysConclusion BC-GP generated accurate identification and detection of resistance markers compared with routine laboratorymethods for Gram-positive organisms in specialized clinical settings providing more rapid results than current routine testing

1 Introduction

Gram-positive bacteria are the most predominant microor-ganisms associated with sepsis in healthcare settings andthe most prevalent cause of bacteremia in patients withhematopoietic stem cell transplantation [1 2] Enterococcalbacteremia is associated with increased risk of mortality

in patients with hematopoietic stem cell transplantationirrespective of susceptibility to vancomycin [3 4] Incardiology units infective endocarditis infectious aneurysmcatheter-related bloodstream infections or surgical site infec-tions after cardiac surgeries are the major infections of whichthe most common causative microorganisms are Gram-positive cocci [5ndash10] In bothmedical units early detection of

HindawiCanadian Journal of Infectious Diseases and Medical MicrobiologyVolume 2017 Article ID 4896791 10 pageshttpsdoiorg10115520174896791

2 Canadian Journal of Infectious Diseases and Medical Microbiology

Gram-positives and resistantmarkers is very critical inmana-ging patient care antibiotic stewardship and preventingspread of resistant microorganisms

As early intervention with antimicrobial therapy is asso-ciated with improved prognosis with each hour of delay asso-ciated with increasedmortality rapid diagnosis is critical [11]The Verigene Gram-positive blood culture assay (BC-GP)(Nanosphere Inc Northbrook IL) is a sample to result mic-roarray system for identification of common Gram-positivebacteria and major resistance markers directly from positiveblood culture Although a number of studies have previouslyevaluated BC-GP reporting performance ranging from 92to 99 agreement with conventional methodology [12ndash16]one limitation of previous reports has been that many of thestudies have been from the United States as well as countriesoutside of Japan with only one published report of limitedscope in Japan [17]

Genetic variation among bacterial lineages circulating indifferent geographical regions of the world can affect thesensitivity of molecular assays based on oligonucleotide pro-bes to detect organisms or resistance markers Studies inHong Kong and Belgium have reported lower BC-GP perfor-mance [18 19]

The purpose of this study was to determine the potentialimpact on patient management and patient outcome of BC-GP in specialized hospitalized settings within the Japanesehealthcare environment which faces a number of challengesAn increasingly aging population and the accompanyingburden of increasing overall healthcare costs have challengedthe healthcare infrastructure Despite methicillin-resistantStaphylococcus aureus (MRSA) accounting for over 90 ofthe hospital-acquired infections caused by resistant bacteriain Japan [20] outsourcing of clinical microbiology testingor no weekend coverage is common in many healthcarefacilities as part of cost-containment This paper representsthe first comprehensive evaluation of BC-GP in Japan tovalidate its clinical performanceThe simulated blood culturestudy includes a library of well-characterized healthcare-associated MRSA (HA-MRSA) community-acquired MRSA(CA-MRSA) and vancomycin-resistant enterococci (VRE)strains circulating in Japan

2 Methods

BC-GPwas evaluated at ToranomonHospital (TH) and Saka-kibara Heart Institute (SHI) in accordance with site-specificinstitutional review board approved study protocols duringJune 26 2012 to March 6 2013 TH is an 1168-bed generalteaching hospital with a 123-bed hematological care unitfor hematopoietic stem cell transplantation performing 140to 160 hematopoietic stem cell transplants per year Themicrobiology laboratory at the hospital operates daily duringday hours SHI is a 320-bed teaching hospital specializing incardiovascular diseases with a caseload of over 1500 open-heart surgeries per year The microbiology laboratory inthe hospital is operated by an outside commercial referencelaboratory The microbiology laboratory operates during theday shift on weekdays and closed on weekends and holidays

Blood cultures were performed at SHI using BacTALERT FA bottles and monitoring with BacTALERT 3D(bioMerieux Marcy lrsquoEtoile France) TH used BACTEC Plusbottles and monitoring with BACTEC 9240 and FX (BectonDickinson Franklin Lakes and NJ) Only one positive bloodculture bottle containing Gram-positive cocci or bacilli perpatient was included in the study Two ml of positive bloodculture medium was stored at minus85C for retesting

Routine microbiological identification and susceptibilitytesting of isolates were performed using conventional identi-fication tests such as bile solubility optochin disk susceptibil-ity and the MicroScanWalkAway system (Beckman CoulterPasadena CA) at TH and the Vitek 2 system (bioMerieuxMarcy lrsquoEtoile France) at SHI Cefoxitin screening for methi-cillin resistance was performed according to CLSI guidelines[21] A latex agglutination test for detection of penicillin-binding protein PBP2a was also utilized [22]

BC-GP testing was performed on positive blood cultureshowing Gram-positive organisms according to the manu-facturerrsquos instructions Briefly a well-mixed 350 120583l sample ofthe blood culture media was pipetted into the sample well ofthe BC-GP nucleic extraction tray placed onto the VerigeneProcessor SP for processing and analysis by the VerigeneReader

An assessmentwas performed to determine the differencein time between reporting of results using BC-GP and cul-ture-based identification and antimicrobial susceptibilityresults for 139 positive blood culture broths For culture-based results the time required until generation of the finalreport was the time between the Gram stain reading andentering of final identification and susceptibility results intothe laboratory information system For BC-GP the time toresult was the time between the Gram stain reading andentering of BC-GP results into the laboratory informationsystem

A challenge set of 208 simulated blood cultureswas const-ructed using type reference strains and clinical strainsfrom different geographical regions in Japan to evaluate BC-GP The clinical strains included organisms that presentedchallenges for commercial identification systems in previousclinical studies at TH and SHI In addition a library ofwell-characterized HA-MRSA CA-MRSA and VRE strainsfrom Japan was also tested Simulated blood culture studieswere performed at Miroku Medical Laboratory (Saku CityNagano Prefecture Japan) Two hundred and eight strainswere adjusted to a turbidity of approximately 100CFUml insterile salineThree hundred 120583l was inoculated into BACTECPlus AerobicF bottles containing 8 to 10ml of human wholeblood (blood type O Tennessee Blood Services MemphisTN) for a final inoculum of 30CFUbottle A BACTEC PlusAnaerobicF bottle was also inoculated for S pneumoniaeand the S anginosus group Each bottle was incubated in theBACTEC system until a positive signal was generated If BC-GP generated negative results 11-fold dilution of the bloodculture medium using sterile distilled water was retested

Each of the positive blood culture isolates at the twohospital sites was stored in 10 skim milk (Difco) at minus85CSpecies identification was confirmed using matrix-assisted

Canadian Journal of Infectious Diseases and Medical Microbiology 3

laser desorption ionization-time of flight mass spectrometry(MALDI-TOF MS) (Microflex LT with Biotyper ver 30software Bruker Daltonik GmbH Bremen Germany) forall strains [23] If the organism was not identified to thespecies level by MALDI-TOFMS (score value lt 20) or iden-tified as Micrococcus Listeria Staphylococcus other than Saureus Streptococcus other than S pyogenes and S agalac-tiae confirmatory testing by PCR-direct sequencing of 16SrDNA or sodA was performed at Juntendo University orTokyo Womenrsquos Medical University [24ndash26] Specific-PCRfor detecting mecA [27] in all staphylococci and vanA vanBin all enterococci was performed [28] The methods usedfor SCCmec typing of community-acquired or healthcare-associated MRSA used to characterized strains have beenpreviously described [29 30]

Concordance was determined in comparison to results ofthe reference methods There was agreement if BC-GP targetdetection agreed with the reference method at either thegenus or the species level The ninety-five percent confidenceintervals (95 CI) and the paired 119905-test were determinedusing GraphPad StatMate (GraphPad Software Inc SanDiego CA)

3 Results

In this study the overall identification agreement betweenBC-GP and the reference method was 327347 (94) forprospective blood cultures at two clinical sites and simulatedblood cultures The combined agreement between PCR andBC-GP for mecA detection was 7173 (97) for prospectiveblood cultures and simulated blood cultures

The combined identification accuracy from both hos-pital sites was 129139 (93) For monomicrobial culturesthe identification accuracy was 121124 (98) Agreementbetween PCR and BC-GP for mecA positivity was 5153(96) Table 1 shows the results from TH Overall 96104(92) organisms were correctly identified by BC-GP to thespecies or genus level including detection of resistance genesAs shown in Table 2 total agreement of BC-GP with thereference method at SHI was 3335 (94) organisms

BC-GP reports the presence of mecA for only S aureusand S epidermidis In this study of the 102 staphylococcalstrains 72 were either S aureus or S epidermidis Discordantresults were due to undetectable mecA S epidermidis organ-isms in polymicrobial cultures containing bothmecA positiveand mecA negative S epidermidis Of the 30 Staphylococcusspp other than S epidermidis and S aureus 21 (70) werepositive formecA including 2 S lugdunensis which could notbe reported asmecA positive by BC-GP

Table 3 shows the difference in time between the gener-ation of BC-GP results and culture-based final identificationand antimicrobial susceptibility results at both hospitals BC-GP results were available at amean of 282 to 510 hours beforeculture-based final identification and susceptibility results atTH At SHI BC-GP generated results at a mean of 345 to1966 hours earlier In comparing the time to final culture-based identification and susceptibility results at TH and SHIwith the exception of S aureus results for S epidermidis andcoagulase-negative staphylococci other than S epidermidis

enterococci and streptococci required significantly (119875 lt005) longer time at SHI (833 1236 1591 and 1991 hoursresp) compared to TH (408 539 361 and 535 hours resp)

Using simulated Gram-positive blood cultures BC-GPcorrectly identified 198208 (95) of the organisms (Table 4)Six streptococci (3) were either incorrectly identified oridentified at the genus level only by BC-GP With respectto the 4 BC-GP false-negative blood culture bottles 1 Spyogenes was correctly identified and 1 S mitis generated apositive Streptococcus genusS pneumoniae signal following11-fold dilution of the blood culture medium The mecAgene was detected in 2020 (100) MRSA organisms rep-resenting community-acquired (SCCmec type IIa IV andV) and healthcare-associated (SCCmec type I IIb III andnontypeable) strains by BC-GP BC-GP detected 1414 (100)vanA and 2020 (100) vanB genes in well-characterizedVRE strains from previous studies in Japan

4 Discussion

Theperformance of BC-GP observed in our studywas similarto previous reports [12ndash16 31ndash33] As clinical microbiologyservices are outsourced or not operating during off-shiftsduring weekdays and closed on weekends and holidaysin many hospitals in Japan rapid diagnostic testing hassignificant potential to impact patient care by dramaticallydecreasing the time to organism identification and antimicro-bial susceptibility results

BC-GP detects mecA in all staphylococci based on mea-surement of signal intensity however reporting is restrictedto S aureus and S epidermidis based on an algorithm inwhichmecA is only reported when S aureus or S epidermidisis detected by BC-GP Future versions of BC-GP shouldconsider modification of the algorithm to allow reporting ofmecA detection for staphylococci other than S aureus or Sepidermidis as 70 of the 30 non-S aureus and S epidermidisstrains in our study were methicillin-resistant Although Sepidermidis is the major coagulase-negative staphylococcalpathogen other staphylococci such as S lugdunensis andS haemolyticus are important pathogens in the healthcareenvironment [34 35]

Polymicrobial positive blood cultures generated themajority of the discordance In contrast the performance ofBC-GP was 121124 (98) in monomicrobial clinical bloodcultures and 198208 (95) in simulated blood culturesIn this study polymicrobial blood cultures accounted for14106 (13) and 333 (9) respectively of the positive bloodcultures at TH and SH Combined polymicrobial culturesrepresented 17139 (12) of the prospective clinical bloodcultures which is consistent with previous studies reporting6 to 20 of all bloodstream infections to be polymicrobial[36ndash38] BC-GP correctly identified all of the organismsin 1217 (70) of polymicrobial cultures In previous BC-GP studies the correct identification rates in polymicrobialcultures ranged from 57 to 86 [14ndash16 31 32] As misleadinginformation can affect clinical diagnosis resulting in inap-propriate selection of antimicrobial agents there is a need tounderstand the limitations of BC-GP


Table1Perfo

rmance

oftheB

C-GPassayatTo

rano

mon

Hospital

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

7873

(94)

4(5)

1(1)

7170

(99)

1(2)

Saureus

1616

(100)

1616

(100)

Methicillin-sensitive

99(100)

99(100)

Methicillin-resistant

77(100)

77(100)

Sepidermidis

3734

(92)

2(5)

1(3)

3434

(100)


21(50)

1(50)a

11(100)

Methicillin-resis

tant

3533

(94)

1(3)

b1(3)

c33

33(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

2422

(92)

2(8)

2019

(95)

1(6)

Scaprae

98(89)

1(11)

76(67)

1(33)

Shominis

88(100)

77(100)

Shaem

olyticu

s4

3(75)

1(25)d

33(100)

Scapitis

11(100)

11(100)

Sschleiferi

11(100)

11(100)

Ssim

ulan

s1

1(100)

11(100)

Streptococcus

97(78)

1(11)

1(11)

65(83)

1(27)

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

11(100)

Sconstellatus

11(100)

e

Other

streptococci

75(72)

1(14)

1(14)

54(80)

Smitis

21(50)

1(50)f

21(50)

1(50)f

Sinfantis

21(50)

1(50)g

11(100)

Stigurinus

22(100)

11(100)

Soralis

11(100)

11(100)

Enterococcus

1716

(94)

1(6)

1616

(100)

Efaecalis

21(50)

1(50)h

11(100)

Vancom

ycin-sensitive

21(50)

1(50)

11(100)

Efaecium

1515

(100)

1515

(100)

Vancom

ycin-sensitive

1515

(100)

1515

(100)

Total

104

96(92)

6(6)

2(2)

9391

(98)

1(1)

1(1)

Other

nontargetG

ram-positives

1510

Bacillus

32

Bsubtilis

21

Bcereus

11

Corynebacterium

128

Cstr

iatum

95

Cjeikeium

33


Table1Con

tinued

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edTo

talisolates

119103

a Polym

icrobialcultu

reof

methicillin-sensitive

andmethicillin-resistant

Sepidermidis

b Polym

icrobialcultu

rewith

Efaecium

c Correctlyidentifi

edas

Staphylococcusbut

notasS

epiderm

idis

SaureusorS

lugdu

nensis

d Polym

icrobialcultu

rewith

Stigurinus

e ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas

ldquocorrectlyidentifi

edrdquofor

each

species

f Identifi

edas

Spn

eumoniae

g Polym

icrobialcultu

rewith

methicillin-resistant

Sepidermidis

h Polym

icrobialcultu

rewith

Escherich

iacoli


Table2Perfo

rmance

oftheB

C-GPassayatSakakibara

HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis


Table 3 Difference in time to final identification and antimicrobial susceptibility report

OrganismDifference in time to resulta

Sakakibara Heart Institute Toranomon HospitalMean (h) Range Mean (h) Range

S aureus 345 (119875 lt 005) 215ndash468 282 (119875 lt 005) 196ndash470S epidermidis 807 (119875 lt 005) 239ndash1609 383 (119875 lt 005) 216ndash725Coagulase-negative staphylococci 1211 (119875 lt 005) 259ndash2174 514 (119875 lt 005) 214ndash722Enterococcus spp 1566 (119875 lt 005) 959ndash2174 336 (119875 lt 005) 234ndash477Streptococcus spp 1966 (119875 lt 005) 423ndash5026 510 (119875 lt 005) 234ndash692aThe difference in time between BC-GP result and final culture-based identification and susceptibility results

An additional concern with polymicrobial cultures is theclinical interpretation of mecA detection and staphylococcaldetection In our 17-polymicrobial cultures 5 samples yielded2 or 3 staphylococcal strains with or without mecA Thismay lead to unnecessary use of vancomycin or underesti-mating infection caused by methicillin-resistant staphylo-cocci Repeating BC-GP on another set of blood culturesmay lessen the risk of this problem For monomicrobialcultures or simulated cultures all of the discrepancies wereobserved with streptococci except for 1 S caprae strain BC-GP misidentification for streptococci included 2 S mitisidentified as S pneumoniae no detection of 2 S pneumoniae2 S anginosus group and 2 S pyogenes Previous reportshave also reported similar results for S mitis S oralis andS pneumoniae [14ndash16 39] As genetic relatedness amongS mitis S oralis and S pneumoniae is well known basedon gt99 homology of 16S rRNA gene sequences [24] BC-GP results for S pneumoniae or Streptococcus with alpha-hemolysiswithout any positive species-specific signals shouldbe carefully interpreted and confirmed by conventionalmethods such as optochin sensitivity or the bile solubilitytest Interestingly 11-fold dilution of the original blood culturemedium can lead to detection of the Streptococcus signal andspecies signal (Table 4) A range of detection depending onthe organism by BC-GP has been reported [40]

The major benefit of utilizing BC-GP is the earlier timefor reporting identification and resistance determinants frompositive blood cultures allowing for earlier selection of appro-priate antimicrobial therapy and implementation of infectioncontrol measures such as isolation and contact precaution[39 41] This has the potential of making a significant impactin the Japanese healthcare delivery system The differencein time between BC-GP results and final culture-basedidentification and susceptibility results shown in Table 3 atTH is consistent with previous reports [14ndash16 18 32 41]On the other hand earlier results of 807 to 1966 hours fororganisms other than S aureus using BC-GP at SHI reflectsthe unavailability of clinical microbiology services duringweekends and holidays As clinical microbiology services areoutsourced in many hospitals in Japan or limited to oneshift during weekdays or not offered during weekends thepotential cost-benefits of retaining blood culture services inhospitals are significant Furthermore training of laboratorypersonnel in the general laboratory to recognize Gram-positive cocci and bacilli will allow BC-GP testing over the

weekend as well as eveningsnights BC-GP provides theopportunity for hospitals to retain in-house a very criticallaboratory service

In conclusion BC-GP provided accurate identificationand detection of resistance markers compared with routineculture-based laboratory methods for Gram-positive organ-isms including CA-MRSA HA-MRSA and VRE strains cir-culating in Japan Minimizing the time to optimizing antimi-crobial therapy using BC-GP may contribute to reducedcosts and improved patient care In 2016 BC-GP receivedregulatory approval in Japan becoming the first multitargetmolecular test for positive blood cultures approved as an invitro diagnostic device to aid in the diagnosis of bacterialbloodstream infections Additional studies will be necessaryto validate the cost-effectiveness of BC-GPwithin the contextof the Japanese healthcare delivery system

Ethical Approval

This study was approved by the TH and SHI internal reviewboards

Competing Interests

All the authors declare no competing interests

Authorsrsquo Contributions

Ken Kikuchi designed and carried out the study and draftedthe manuscript Mari Matsuda Shigekazu Iguchi TomonoriMizutani Kaori Sansaka Kenta Negishi Kimie ShimadaShigeyuki Notake Hideji Yanagisawa and Reiko Yabusakicarried out the laboratory works Keiichi Hiramatsu MichiruTega-Ishii JunUmemura Hiroshi Takahashi Hideki Araokaand Akiko Yoneyama supervised the data collection andcoordinated and participated in designing the study

Acknowledgments

This study was supported in part by a Grant-in-Aid(S0991013) from the Ministry of Education Culture Sport


Table 4 Detection of Gram-positive bacteria and resistance genes in simulated blood cultures by BC-GP

Organism Total no of strains No () of isolatescorrectly identified not detected incorrectly identified

Staphylococcus 54 54 (100)S aureus 35 35 (100)

Methicillin-sensitivemecAminus 15 15 (100)Methicillin-resistantmecA+ 20 20 (100)Health-care associated 10 10 (100)Community acquired 10 10 (100)

S epidermidis 1 1 (100)Methicillin-sensitivemecAminus 1 1 (100)

S lugdunensis 8 8 (100)Other CNS 10 10 (100)

S hominis 2 2 (100)S haemolyticus 2 2 (100)S saprophyticus 2 2 (100)S capitis 1 1 (100)S cohnii 1 1 (100)S warneri 1 1 (100)S pseudintermedius 1 1 (100)

Streptococcus 87 77 (88) 4 (5) 6 (7)S pyogenes 9 8 (89) 1 (11)a

S agalactiae 8 8 (100)S dysgalactiae 8 8 (100)S anginosus groupb 10 8 (80) 2 (20)S anginosus 3 2 (66) 1 (34)c

S constellatus 4 3 (75) 1 (25)c

S intermedius 3 3 (100)S pneumoniae 22 20 (91) 2 (9)d

Other streptococci 30 25 (86) 3 (10) 2 (7)S mitis 12 9 (75) 1 (8) 2 (17)e

S oralis 4 4 (100)S infantis 2 2 (100)S mutans 2 1 (50) 1 (50)S sobrinus 1 0 (0) 1 (100)S sanguinis 1 1 (100)

S parasanguinis 1 1 (100)S peroris 1 1 (100)S australis 1 1 (100)S tigurinus 1 1 (100)S cristatus 1 1 (100)S gordonii 1 1 (100)S gallolyticus 1 1 (100)S lutetiensis 1 1 (100)

Enterococcus 57 57 (100)E faecalis 32 32 (100)

Vancomycin-sensitive 18 18 (100)Vancomycin-resistant vanA+ 4 4 (100)Vancomycin-resistant vanB+ 10 10 (100)

E faecium 25 25 (100)Vancomycin-sensitive 5 5 (100)Vancomycin-resistant vanA+ 10 10 (100)Vancomycin-resistant vanB+ 10 10 (100)

Listeria spp 5 5 (100)


Table 4 Continued


Micrococcus spp 5 5 (100)Total 208 198 (95) 4 (2) 6 (3)aNot detected initially but positive using 11-fold diluted blood culture samplebStreptococcus sp belonging to the S anginosus group is identified as S anginosus group by BC-GPcPositive signal for Streptococcus but no signal for S anginosus groupdPositive signal for Streptococcus but no signal for S pneumoniaeeMisidentified as S pneumoniae

Science and Technology Japan (MEXT) for the Foundationof Strategic Research Projects in Private Universities

References

[1] P Cappellano C Viscoli P Bruzzi M T Van Lint C A PPereira and A Bacigalupo ldquoEpidemiology and risk factors forbloodstream infections after allogeneic hematopoietic stem celltransplantionrdquo New Microbiologica vol 30 no 2 pp 89ndash1002007

[2] M Mikulska V Del Bono P Bruzzi et al ldquoMortality afterbloodstream infections in allogeneic haematopoietic stem celltransplant (HSCT) recipientsrdquo Infection vol 40 no 3 pp 271ndash278 2012

[3] M Mikulska V Del Bono R Prinapori et al ldquoRisk factors forenterococcal bacteremia in allogeneic hematopoietic stem celltransplant recipientsrdquo Transplant Infectious Disease vol 12 no6 pp 505ndash512 2010

[4] J Vydra R M Shanley I George et al ldquoEnterococcal bac-teremia is associated with increased risk of mortality in recip-ients of allogeneic hematopoietic stem cell transplantationrdquoClinical Infectious Diseases vol 55 no 6 pp 764ndash770 2012

[5] T J Cahill and B D Prendergast ldquoInfective endocarditisrdquo TheLancet vol 387 no 10021 pp 882ndash893 2016

[6] A Michalopoulos S Geroulanos E S Rosmarakis and M EFalagas ldquoFrequency characteristics and predictors of micro-biologically documented nosocomial infections after cardiacsurgeryrdquo European Journal of Cardio-Thoracic Surgery vol 29no 4 pp 456ndash460 2006

[7] G S Oderich J M Panneton T C Bower et al ldquoInfectedaortic aneurysms aggressive presentation complicated earlyoutcome but durable resultsrdquo Journal of Vascular Surgery vol34 no 5 pp 900ndash908 2001

[8] C D Owens and K Stoessel ldquoSurgical site infections epi-demiology microbiology and preventionrdquo Journal of HospitalInfection vol 70 supplement 2 pp 3ndash10 2008

[9] B O Soderquist ldquoSurgical site infections in cardiac surgerymicrobiologyrdquo APMIS vol 115 no 9 pp 1008ndash1011 2007

[10] J Wojkowska-Mach M Baran R Drwiła et al ldquoFactors influ-encing the occurence of nosocomial bloodstream infectionsobserved in thoracic and cardiosurgical postoperative careunitsrdquo Anaesthesiology intensive therapy vol 44 no 1 pp 16ndash20 2012

[11] A Kumar D Roberts B Light et al ldquoDuration of hypotensionbefore initiation of effective antimicrobial therapy is the criticaldeterminant of survival in human septic shockrdquo Critical CareMedicine vol 34 no 6 pp 1589ndash1596 2006

[12] B W Buchan C C Ginocchio R Manii et al ldquoMultiplex iden-tification of gram-positive bacteria and resistance determinants

directly from positive blood culture broths evaluation of anautomated microarray-based nucleic acid testrdquo PLoS Medicinevol 10 Article ID e1001478 2013

[13] J Mestas C M Polanco S Felsenstein and J D BardldquoPerformance of the verigene gram-positive blood culture assayfor direct detection of gram-positive organisms and resistancemarkers in a pediatric hospitalrdquo Journal of ClinicalMicrobiologyvol 52 no 1 pp 283ndash287 2014

[14] L P Samuel R J Tibbetts A Agotesku M Fey R Hensley andF A Meier ldquoEvaluation of a microarray-based assay for rapididentification of gram-positive organisms and resistance mark-ers in positive blood culturesrdquo Journal of Clinical Microbiologyvol 51 no 4 pp 1188ndash1192 2013

[15] K V Sullivan N N Turner S S Roundtree et al ldquoRapid det-ection of gram-positive organisms by use of the verigene gram-positive blood culture nucleic acid test and the bactalertpediatric fan system in a multicenter pediatric evaluationrdquoJournal of Clinical Microbiology vol 51 no 11 pp 3579ndash35842013

[16] C M Wojewoda L Sercia M Navas et al ldquoEvaluation of theverigene gram-positive blood culture nucleic acid test for rapiddetection of bacteria and resistance determinantsrdquo Journal ofClinical Microbiology vol 51 no 7 pp 2072ndash2076 2013

[17] H Suzuki S Hitomi Y Yaguchi et al ldquoProspective interven-tion study with a microarray-based multiplexed automatedmolecular diagnosis instrument (Verigene system) for the rapiddiagnosis of bloodstream infections and its impact on theclinical outcomesrdquo Journal of Infection and Chemotherapy vol21 no 12 pp 849ndash856 2015

[18] M Dodemont R De Mendonca C Nonhoff S Roisin andO Denis ldquoEvaluation of Verigene Gram-positive blood cultureassay performance for bacteremic patientsrdquo European Journal ofClinical Microbiology and Infectious Diseases vol 34 no 3 pp473ndash477 2014

[19] G K H Siu J H K Chen T K Ng et al ldquoPerformance evalua-tion of the verigene gram-positive and gram-negative bloodculture test for direct identification of bacteria and theirresistance determinants from positive blood cultures in HongKongrdquo PLOS ONE vol 10 no 10 Article ID e0139728 2015

[20] Japan Nosocomial Infections Surveillance (JANIS) ldquoOpenReport 2013-Hospital-acquired Infection Surveillancerdquo 2014httpswwwnih-janisjpenglishreportopen report201331ken Open Report Eng 201300pdf

[21] Clinical and Laboratory Standards Institute Performance Stan-dards for Antimicrobial Susceptibility Testing Twenty-FirstInformational Supplement M100-S21 Clinical and LaboratoryStandards Institute Wayne Pa USA 2011


[22] Y Nakatomi and J Sugiyama ldquoA rapid latex agglutination assayfor the detection of penicillin-binding protein 21015840rdquoMicrobiologyand Immunology vol 42 no 11 pp 739ndash743 1998

[23] N Matsuda M Matsuda S Notake et al ldquoEvaluation of asimple protein extraction method for species identification ofclinically relevant staphylococci by matrix-assisted laser des-orption ionizationmdashtime of flight mass spectrometryrdquo Journalof Clinical Microbiology vol 50 no 12 pp 3862ndash3866 2012

[24] Y Kawamura X-G Hou F Sultana H Miura and TEzaki ldquoDetermination of 16S rRNA sequences of Streptococcusmitis and Streptococcus gordonii and phylogenetic relationshipsamong members of the genus Streptococcusrdquo InternationalJournal of Systematic Bacteriology vol 45 no 2 pp 406ndash4081995

[25] Y Kawamura R AWhiley S-E Shu T Ezaki and JMHardieldquoGenetic approaches to the identification of the mitis groupwithin the genus StreptococcusrdquoMicrobiology vol 145 no 9 pp2605ndash2613 1999

[26] C Poyart G Quesne C Boumaila and P Trieu-Cuot ldquoRapidand accurate species-level identification of coagulase-negativestaphylococci by using the sodA gene as a targetrdquo Journal ofClinical Microbiology vol 39 no 12 pp 4296ndash4301 2001

[27] K Murakami W Minamide K Wada E Nakamura H Tera-oka and S Watanabe ldquoIdentification of methicillin-resistantstrains of staphylococci by polymerase chain reactionrdquo Journalof Clinical Microbiology vol 29 no 10 pp 2240ndash2244 1991

[28] S Dutka-Malen S Evers and P Courvalin ldquoDetection ofglycopeptide resistance genotypes and identification to thespecies level of clinically relevant enterococci by PCRrdquo Journalof Clinical Microbiology vol 33 no 1 pp 24ndash27 1995

[29] Y Kondo T Ito X X Ma et al ldquoCombination of multiplexPCRs for staphylococcal cassette chromosomemec type assign-ment rapid identification system for mec ccr and major diff-erences in junkyard regionsrdquo Antimicrobial Agents and Chemo-therapy vol 51 no 1 pp 264ndash274 2007

[30] C Piao T Karasawa K Totsuka T Uchiyama and K KikuchildquoProspective surveillance of community-onset and healthcare-associated methicillin-resistant Staphylococcus aureus isolatedfrom a university-affiliated hospital in JapanrdquoMicrobiology andImmunology vol 49 no 11 pp 959ndash970 2005

[31] S L Aitken V S Hemmige H L Koo N N Vuong T MLasco and K W Garey ldquoReal-world performance of a micro-array-based rapid diagnostic for Gram-positive bloodstreaminfections and potential utility for antimicrobial stewardshiprdquoDiagnostic Microbiology and Infectious Disease vol 81 no 1 pp4ndash8 2015

[32] S G Beal J Ciurca G Smith et al ldquoEvaluation of the nanosph-ere verigene gram-positive blood culture assay with the ver-saTREKblood culture systemand assessment of possible impacton selected patientsrdquo Journal of ClinicalMicrobiology vol 51 no12 pp 3988ndash3992 2013

[33] R M Martinez E R Bauerle F C Fang and S M Butler-Wu ldquoEvaluation of three rapid diagnostic methods for directidentification of microorganisms in positive blood culturesrdquoJournal of Clinical Microbiology vol 52 no 7 pp 2521ndash25292014

[34] K L Frank J L Del Pozo and R Patel ldquoFrom clinical micro-biology to infection pathogenesis how daring to be differentworks for staphylococcus lugdunensisrdquo Clinical MicrobiologyReviews vol 21 no 1 pp 111ndash133 2008

[35] E M Barros H Ceotto M C F Bastos K R N Dos Santosand M Giambiagi-deMarval ldquoStaphylococcus haemolyticus as

an important hospital pathogen and carrier of methicillinresistance genesrdquo Journal of Clinical Microbiology vol 50 no1 pp 166ndash168 2012

[36] G S Cooper D S Havlir D M Shlaes and R A SalataldquoPolymicrobial bacteremia in the late 1980s predictors ofoutcome and review of the literaturerdquo Medicine vol 69 no 2pp 114ndash123 1990

[37] S Sancho A Artero R Zaragoza J J Camarena R Gonzalezand J M Nogueira ldquoImpact of nosocomial polymicrobialbloodstream infections on the outcome in critically ill patientsrdquoEuropean Journal of Clinical Microbiology and Infectious Dis-eases vol 31 no 8 pp 1791ndash1796 2012

[38] S Y Park K-H Park K M Bang et al ldquoClinical significanceand outcome of polymicrobial Staphylococcus aureus bac-teremiardquoThe Journal of Infection vol 65 no 2 pp 119ndash127 2012

[39] K Alby L M Daniels D J Weber and M B Miller ldquoDevelop-ment of a treatment algorithm for streptococci and enterococcifrom positive blood cultures identified with the Verigene gram-positive blood culture assayrdquo Journal of Clinical Microbiologyvol 51 no 11 pp 3869ndash3871 2013

[40] T Raich and S Powell ldquoIdentification of bacterial and fungalpathogens from positive blood culture bottles a microarray-based approachrdquo Methods in Molecular Biology vol 1237 pp73ndash90 2015

[41] D G Roshdy A Tran N LeCroy et al ldquoImpact of a rapid mic-roarray-based assay for identification of positive blood culturesfor treatment optimization for patients with streptococcal andenterococcal bacteremiardquo Journal of Clinical Microbiology vol53 no 4 pp 1411ndash1414 2015

Submit your manuscripts athttpswwwhindawicom

Stem CellsInternational

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014


MEDIATORSINFLAMMATION

of


Behavioural Neurology

EndocrinologyInternational Journal of



Disease Markers


BioMed Research International

OncologyJournal of



Oxidative Medicine and Cellular Longevity


PPAR Research

The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014

Immunology ResearchHindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Journal of

ObesityJournal of



Computational and Mathematical Methods in Medicine

OphthalmologyJournal of


Diabetes ResearchJournal of



Research and TreatmentAIDS


Gastroenterology Research and Practice


Parkinsonrsquos Disease

Evidence-Based Complementary and Alternative Medicine

Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


Gram-positives and resistantmarkers is very critical inmana-ging patient care antibiotic stewardship and preventingspread of resistant microorganisms

As early intervention with antimicrobial therapy is asso-ciated with improved prognosis with each hour of delay asso-ciated with increasedmortality rapid diagnosis is critical [11]The Verigene Gram-positive blood culture assay (BC-GP)(Nanosphere Inc Northbrook IL) is a sample to result mic-roarray system for identification of common Gram-positivebacteria and major resistance markers directly from positiveblood culture Although a number of studies have previouslyevaluated BC-GP reporting performance ranging from 92to 99 agreement with conventional methodology [12ndash16]one limitation of previous reports has been that many of thestudies have been from the United States as well as countriesoutside of Japan with only one published report of limitedscope in Japan [17]

Genetic variation among bacterial lineages circulating indifferent geographical regions of the world can affect thesensitivity of molecular assays based on oligonucleotide pro-bes to detect organisms or resistance markers Studies inHong Kong and Belgium have reported lower BC-GP perfor-mance [18 19]

The purpose of this study was to determine the potentialimpact on patient management and patient outcome of BC-GP in specialized hospitalized settings within the Japanesehealthcare environment which faces a number of challengesAn increasingly aging population and the accompanyingburden of increasing overall healthcare costs have challengedthe healthcare infrastructure Despite methicillin-resistantStaphylococcus aureus (MRSA) accounting for over 90 ofthe hospital-acquired infections caused by resistant bacteriain Japan [20] outsourcing of clinical microbiology testingor no weekend coverage is common in many healthcarefacilities as part of cost-containment This paper representsthe first comprehensive evaluation of BC-GP in Japan tovalidate its clinical performanceThe simulated blood culturestudy includes a library of well-characterized healthcare-associated MRSA (HA-MRSA) community-acquired MRSA(CA-MRSA) and vancomycin-resistant enterococci (VRE)strains circulating in Japan

2 Methods

BC-GPwas evaluated at ToranomonHospital (TH) and Saka-kibara Heart Institute (SHI) in accordance with site-specificinstitutional review board approved study protocols duringJune 26 2012 to March 6 2013 TH is an 1168-bed generalteaching hospital with a 123-bed hematological care unitfor hematopoietic stem cell transplantation performing 140to 160 hematopoietic stem cell transplants per year Themicrobiology laboratory at the hospital operates daily duringday hours SHI is a 320-bed teaching hospital specializing incardiovascular diseases with a caseload of over 1500 open-heart surgeries per year The microbiology laboratory inthe hospital is operated by an outside commercial referencelaboratory The microbiology laboratory operates during theday shift on weekdays and closed on weekends and holidays

Blood cultures were performed at SHI using BacTALERT FA bottles and monitoring with BacTALERT 3D(bioMerieux Marcy lrsquoEtoile France) TH used BACTEC Plusbottles and monitoring with BACTEC 9240 and FX (BectonDickinson Franklin Lakes and NJ) Only one positive bloodculture bottle containing Gram-positive cocci or bacilli perpatient was included in the study Two ml of positive bloodculture medium was stored at minus85C for retesting

Routine microbiological identification and susceptibilitytesting of isolates were performed using conventional identi-fication tests such as bile solubility optochin disk susceptibil-ity and the MicroScanWalkAway system (Beckman CoulterPasadena CA) at TH and the Vitek 2 system (bioMerieuxMarcy lrsquoEtoile France) at SHI Cefoxitin screening for methi-cillin resistance was performed according to CLSI guidelines[21] A latex agglutination test for detection of penicillin-binding protein PBP2a was also utilized [22]

BC-GP testing was performed on positive blood cultureshowing Gram-positive organisms according to the manu-facturerrsquos instructions Briefly a well-mixed 350 120583l sample ofthe blood culture media was pipetted into the sample well ofthe BC-GP nucleic extraction tray placed onto the VerigeneProcessor SP for processing and analysis by the VerigeneReader

An assessmentwas performed to determine the differencein time between reporting of results using BC-GP and cul-ture-based identification and antimicrobial susceptibilityresults for 139 positive blood culture broths For culture-based results the time required until generation of the finalreport was the time between the Gram stain reading andentering of final identification and susceptibility results intothe laboratory information system For BC-GP the time toresult was the time between the Gram stain reading andentering of BC-GP results into the laboratory informationsystem

A challenge set of 208 simulated blood cultureswas const-ructed using type reference strains and clinical strainsfrom different geographical regions in Japan to evaluate BC-GP The clinical strains included organisms that presentedchallenges for commercial identification systems in previousclinical studies at TH and SHI In addition a library ofwell-characterized HA-MRSA CA-MRSA and VRE strainsfrom Japan was also tested Simulated blood culture studieswere performed at Miroku Medical Laboratory (Saku CityNagano Prefecture Japan) Two hundred and eight strainswere adjusted to a turbidity of approximately 100CFUml insterile salineThree hundred 120583l was inoculated into BACTECPlus AerobicF bottles containing 8 to 10ml of human wholeblood (blood type O Tennessee Blood Services MemphisTN) for a final inoculum of 30CFUbottle A BACTEC PlusAnaerobicF bottle was also inoculated for S pneumoniaeand the S anginosus group Each bottle was incubated in theBACTEC system until a positive signal was generated If BC-GP generated negative results 11-fold dilution of the bloodculture medium using sterile distilled water was retested

Each of the positive blood culture isolates at the twohospital sites was stored in 10 skim milk (Difco) at minus85CSpecies identification was confirmed using matrix-assisted




3 Results







4 Discussion





Table1Perfo

rmance

oftheB

C-GPassayatTo

rano

mon

Hospital

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

7873

(94)

4(5)

1(1)

7170

(99)

1(2)

Saureus

1616

(100)

1616

(100)


99(100)

99(100)


77(100)

77(100)

Sepidermidis

3734

(92)

2(5)

1(3)

3434

(100)


21(50)

1(50)a

11(100)

Methicillin-resis

tant

3533

(94)

1(3)

b1(3)

c33

33(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

2422

(92)

2(8)

2019

(95)

1(6)

Scaprae

98(89)

1(11)

76(67)

1(33)

Shominis

88(100)

77(100)

Shaem

olyticu

s4

3(75)

1(25)d

33(100)

Scapitis

11(100)

11(100)

Sschleiferi

11(100)

11(100)

Ssim

ulan

s1

1(100)

11(100)

Streptococcus

97(78)

1(11)

1(11)

65(83)

1(27)

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

11(100)

Sconstellatus

11(100)

e

Other

streptococci

75(72)

1(14)

1(14)

54(80)

Smitis

21(50)

1(50)f

21(50)

1(50)f

Sinfantis

21(50)

1(50)g

11(100)

Stigurinus

22(100)

11(100)

Soralis

11(100)

11(100)

Enterococcus

1716

(94)

1(6)

1616

(100)

Efaecalis

21(50)

1(50)h

11(100)

Vancom

ycin-sensitive

21(50)

1(50)

11(100)

Efaecium

1515

(100)

1515

(100)

Vancom

ycin-sensitive

1515

(100)

1515

(100)

Total

104

96(92)

6(6)

2(2)

9391

(98)

1(1)

1(1)

Other

nontargetG

ram-positives

1510

Bacillus

32

Bsubtilis

21

Bcereus

11

Corynebacterium

128

Cstr

iatum

95

Cjeikeium

33


Table1Con

tinued

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edTo

talisolates

119103

a Polym

icrobialcultu

reof



Sepidermidis

b Polym

icrobialcultu

rewith

Efaecium

c Correctlyidentifi

edas

Staphylococcusbut

notasS

epiderm

idis

SaureusorS

lugdu

nensis

d Polym

icrobialcultu

rewith

Stigurinus

e ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

f Identifi

edas

Spn

eumoniae

g Polym

icrobialcultu

rewith


Sepidermidis

h Polym

icrobialcultu

rewith

Escherich

iacoli


Table2Perfo

rmance

oftheB


HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis










Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of



















3 Results







4 Discussion





Table1Perfo

rmance

oftheB

C-GPassayatTo

rano

mon

Hospital

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

7873

(94)

4(5)

1(1)

7170

(99)

1(2)

Saureus

1616

(100)

1616

(100)


99(100)

99(100)


77(100)

77(100)

Sepidermidis

3734

(92)

2(5)

1(3)

3434

(100)


21(50)

1(50)a

11(100)

Methicillin-resis

tant

3533

(94)

1(3)

b1(3)

c33

33(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

2422

(92)

2(8)

2019

(95)

1(6)

Scaprae

98(89)

1(11)

76(67)

1(33)

Shominis

88(100)

77(100)

Shaem

olyticu

s4

3(75)

1(25)d

33(100)

Scapitis

11(100)

11(100)

Sschleiferi

11(100)

11(100)

Ssim

ulan

s1

1(100)

11(100)

Streptococcus

97(78)

1(11)

1(11)

65(83)

1(27)

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

11(100)

Sconstellatus

11(100)

e

Other

streptococci

75(72)

1(14)

1(14)

54(80)

Smitis

21(50)

1(50)f

21(50)

1(50)f

Sinfantis

21(50)

1(50)g

11(100)

Stigurinus

22(100)

11(100)

Soralis

11(100)

11(100)

Enterococcus

1716

(94)

1(6)

1616

(100)

Efaecalis

21(50)

1(50)h

11(100)

Vancom

ycin-sensitive

21(50)

1(50)

11(100)

Efaecium

1515

(100)

1515

(100)

Vancom

ycin-sensitive

1515

(100)

1515

(100)

Total

104

96(92)

6(6)

2(2)

9391

(98)

1(1)

1(1)

Other

nontargetG

ram-positives

1510

Bacillus

32

Bsubtilis

21

Bcereus

11

Corynebacterium

128

Cstr

iatum

95

Cjeikeium

33


Table1Con

tinued

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edTo

talisolates

119103

a Polym

icrobialcultu

reof



Sepidermidis

b Polym

icrobialcultu

rewith

Efaecium

c Correctlyidentifi

edas

Staphylococcusbut

notasS

epiderm

idis

SaureusorS

lugdu

nensis

d Polym

icrobialcultu

rewith

Stigurinus

e ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

f Identifi

edas

Spn

eumoniae

g Polym

icrobialcultu

rewith


Sepidermidis

h Polym

icrobialcultu

rewith

Escherich

iacoli


Table2Perfo

rmance

oftheB


HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis










Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table1Perfo

rmance

oftheB

C-GPassayatTo

rano

mon

Hospital

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

7873

(94)

4(5)

1(1)

7170

(99)

1(2)

Saureus

1616

(100)

1616

(100)


99(100)

99(100)


77(100)

77(100)

Sepidermidis

3734

(92)

2(5)

1(3)

3434

(100)


21(50)

1(50)a

11(100)

Methicillin-resis

tant

3533

(94)

1(3)

b1(3)

c33

33(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

2422

(92)

2(8)

2019

(95)

1(6)

Scaprae

98(89)

1(11)

76(67)

1(33)

Shominis

88(100)

77(100)

Shaem

olyticu

s4

3(75)

1(25)d

33(100)

Scapitis

11(100)

11(100)

Sschleiferi

11(100)

11(100)

Ssim

ulan

s1

1(100)

11(100)

Streptococcus

97(78)

1(11)

1(11)

65(83)

1(27)

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

11(100)

Sconstellatus

11(100)

e

Other

streptococci

75(72)

1(14)

1(14)

54(80)

Smitis

21(50)

1(50)f

21(50)

1(50)f

Sinfantis

21(50)

1(50)g

11(100)

Stigurinus

22(100)

11(100)

Soralis

11(100)

11(100)

Enterococcus

1716

(94)

1(6)

1616

(100)

Efaecalis

21(50)

1(50)h

11(100)

Vancom

ycin-sensitive

21(50)

1(50)

11(100)

Efaecium

1515

(100)

1515

(100)

Vancom

ycin-sensitive

1515

(100)

1515

(100)

Total

104

96(92)

6(6)

2(2)

9391

(98)

1(1)

1(1)

Other

nontargetG

ram-positives

1510

Bacillus

32

Bsubtilis

21

Bcereus

11

Corynebacterium

128

Cstr

iatum

95

Cjeikeium

33


Table1Con

tinued

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edTo

talisolates

119103

a Polym

icrobialcultu

reof



Sepidermidis

b Polym

icrobialcultu

rewith

Efaecium

c Correctlyidentifi

edas

Staphylococcusbut

notasS

epiderm

idis

SaureusorS

lugdu

nensis

d Polym

icrobialcultu

rewith

Stigurinus

e ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

f Identifi

edas

Spn

eumoniae

g Polym

icrobialcultu

rewith


Sepidermidis

h Polym

icrobialcultu

rewith

Escherich

iacoli


Table2Perfo

rmance

oftheB


HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis










Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table1Con

tinued

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edTo

talisolates

119103

a Polym

icrobialcultu

reof



Sepidermidis

b Polym

icrobialcultu

rewith

Efaecium

c Correctlyidentifi

edas

Staphylococcusbut

notasS

epiderm

idis

SaureusorS

lugdu

nensis

d Polym

icrobialcultu

rewith

Stigurinus

e ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

f Identifi

edas

Spn

eumoniae

g Polym

icrobialcultu

rewith


Sepidermidis

h Polym

icrobialcultu

rewith

Escherich

iacoli


Table2Perfo

rmance

oftheB


HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis










Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Table2Perfo

rmance

oftheB


HeartInstitute

Organism

BC-G

P(to

tal)

BC-G

P(m

onom

icrobialcultu

res)

Num

bero

forganism

sNum

ber()o

fisolates

Num

bero

forganism

sNum

ber()o

fisolates

Correctlyidentifi

edNot

detected

Incorrectly

identifi

edCorrectlyidentifi

edNot

detected

Incorrectly

identifi

edStaphylococcus

2424

(100)

2222

(100)

Saureus

77(100)

77(100)


66(100)

66(100)


11(100)

11(100)

Sepidermidis

1212

(100)

1111(100)


22(100)

22(100)


1010

(100)

99(100)

Slugdun

ensis

11(100)

11(100)

Other

CNS

44(100)

33(100)

Shominis

22(100)

22(100)

Shaem

olyticu

s1

1(100)

Scapitis

11(100)

11(100)

Streptococcus

88(100)

87(87)

1(13)b

Spyogenes

10(0)

1(100)

a1

0(0)

1(100)

b

Sagalactia

e1

1(100)

11(100)

Sanginosusg

roup

22(100)

22(100)

Sanginosus

22(100)b

22(100)c

Other

streptococci

44(100)

44(100)

Soralis

22(100)

22(100)

Ssanguinis

11(100)

11(100)

Sparasanguinis

11(100)

11(100)

Enterococcus

21(50)

1(50)

Efaecalis

10(0)

1(100)

c

Vancom

ycin-sensitive

10(0)

1(100)

c

Efaecium

11(100)

Vancom

ycin-sensitive

11(100)

Liste

riaspp

11(100)

11(100)

Total

3533

(94)

1(3)

1(3)

3130

(97)

1(3)

Other

nontargetG

ram-positives

11

Corynebacterium

striatum

11

Totalisolates

3632

a Identified

totheg

enus

levelbu

tnot

tothes

pecies

level

b ldquoSanginosusg

roup

rdquoidentified

bytheB

C-GPassayisdefin

edas


edrdquofor

each

species

c Polym

icrobialcultu

rewith

Sepidermidis










Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

























Ethical Approval


Competing Interests




Acknowledgments






















Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of




































Table 4 Continued




References


















































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of











































of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















potential impact of rapid blood culture testing for gram-positive … · 2019. 7. 30. · rdna or...

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