Fluorescence in situ hybridisation (FISH) accelerates identification of Gram-positive cocci in positive blood cultures

Gescher, Dorothee Maria; Kovacevic, Dragoljub; Schmiedel, Dinah; Siemoneit, S.; Mallmann, Christian; Halle, E.; Göbel, Ulf B.; Moter, Annette

doi:10.1016/j.ijantimicag.2008.06.007

Cited by 58 publications

(37 citation statements)

References 33 publications

(26 reference statements)

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“…However, these expensive amplification techniques will probably not be used when starting from positive blood culture bottles. Indeed, growth in blood culture bottles provides sufficiently large amounts of bacteria for identification without molecular amplification, such as the use of fluorescent probes (fluorescence in situ hybridization) [52] or the bacterial identification by MALDI-TOF mass spectrometry (MS) [53]. MALDI-TOF MS has revolutionized clinical microbiology by allowing a rapid, accurate, and cheap identification of bacteria, including GBS [54].…”

Section: Detection Of Pathogens In Positive Blood Culturementioning

confidence: 99%

Molecular-based Screening for Perinatal Group B Streptococcal Infection: Implications for Prevention and Therapy

2013

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Group B streptococci (GBS) are a leading cause of infectious neonatal morbidity and mortality. Timely and accurate identification of colonized pregnant women is imperative to implement intrapartum antibioprophylaxis (IAP) to reduce the risk of early neonatal sepsis. Current guidelines recommend screening for GBS carriage with vaginal-rectal cultures. However, cultures require 24-72 h, thus precluding their use for intrapartum screening and these are only performed at 35-37 weeks gestation. New rapid molecular-based tests can detect GBS within hours. They have the potential to be used intrapartum and to allow for selective IAP in women carrying GBS. An advantage is that they can sometimes be performed by non-laboratory staff in the labor suite, thus avoiding delays in sample transfers to the microbiology laboratory. Another possible use of molecular-based assays is for the diagnosis of neonatal sepsis, where tests with a short turnaround time and high sensitivity and specificity are crucial. In this situation, the detection of microorganisms once antibiotic therapy has already been started is important, as treatment is started immediately once sepsis is suspected without waiting for microbiological confirmation. In this article, we discuss the state-of-the-art molecular-based tests available for GBS screening during pregnancy, as well as their implications for IAP for the diagnosis and prevention of neonatal sepsis.

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Section: Detection Of Pathogens In Positive Blood Culturementioning

confidence: 99%

Molecular-based Screening for Perinatal Group B Streptococcal Infection: Implications for Prevention and Therapy

2013

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“…Gescher and colleagues designed a panel of FISH probes that were genus-specific and species-specific for Gram-positive cocci (staphylococci, streptococci, enterococci and others) to identify organisms from blood cultures. This FISH assay had an overall sensitivity of 98.7%, specificity of 99% and achieved rapid and reliable detection of Gram-positive cocci from clinical blood culture specimens [12]. Kudo and colleagues used FISH to identify microorganisms in blood cultures of 60 patients with suspected sepsis [13].…”

Section: Hybridization-based Fish Techniquesmentioning

confidence: 99%

Molecular microbiological methods in the diagnosis of neonatal sepsis

Venkatesh

Flores

Luna

et al. 2010

Expert Review of Anti-infective Therapy

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Neonatal sepsis is a major cause of neonatal mortality and morbidity. The current gold standard for diagnosis of sepsis, namely blood culture, suffers from low sensitivity and a reporting delay of approximately 48-72 h. Rapid detection of sepsis and institution of antimicrobial therapy may improve patient outcomes. Rapid and sensitive tests that can inform clinicians regarding the institution or optimization of antimicrobial therapy are urgently needed. The ideal diagnostic test should have adequate specificity and negative predictive value to reliably exclude sepsis and avoid unnecessary antibiotic therapy. We comprehensively searched for neonatal studies that evaluated molecular methods for diagnosis of sepsis. We identified 19 studies that were assessed with respect to assay methodology and diagnostic characteristics. In addition, we also reviewed newer molecular microbiological assays of relevance that have not been fully evaluated in neonates. Molecular methods offer distinct advantages over blood cultures, including increased sensitivity and rapid diagnosis. However, diagnostic accuracy and cost-effectiveness should be established before implementation in clinical practice. Keywords diagnosis; FISH; microarray; molecular; neonate; PCR; sepsis Neonatal sepsis is a frequent life-threatening problem in neonatal intensive care units, particularly in very-low-birthweight infants (VLBW; infants with birthweight <1500 g) [1][2][3]. Early-onset sepsis (EOS; sepsis in infants <72 h old) occurs in 1.5-1.9% of VLBW infants, and late-onset sepsis (LOS; onset after 72 h of life) in approximately 20% [1]. Coagulasenegative staphylococci (CONS), Staphylococcus aureus and fungi are responsible for most neonatal infections [1]. Neonatal mortality in LOS is approximately 18% overall and 36% in Gram-negative sepsis. Sepsis also increases neonatal morbidities including patent ductus arteriosus, need for intravascular access, need for parenteral nutrition, bronchopulmonary † Author for correspondence: Tel.: +1 832 824 3206, Fax: +1 832 825 3204, mohanv@bcm.edu.For reprint orders, please contact reprints@expert-reviews.com Financial & competing interests disclosureFunding for color printing of the figures was provided by AdvanDx (MA, USA). Mohan Pammi Venkatesh is funded by CHRCDA grant (NIH K12 HD41648). James Versalovic is supported by R01 AT004326-01A1 NIH/NCCAM, R01 DK065075-01 NIH/NIDDK and 1UH2HG083990-01 NIH/NHGRI. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed. No writing assistance was utilized in the production of this manuscript. NIH Public Access Author ManuscriptExpert Rev Anti Infect Ther. Author manuscript; available in PMC 2011 July 1. . In a large cohort of 6093 extremely low birthweight infants (ELBW; birthweight ≤1000 g), infected infants had a significantly higher incidence of adverse developmental outc...

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“…The most recent generation of automates can detect even small bacterial growth [6]–[8]. When growth is detected by the automate, it is possible to perform direct identification of bacteria by molecular biology, such as universal amplification and sequencing [9], nucleic acid-based fluorescence hybridisation probes, such as FISH [10]–[12], DNA microarrays [13] or molecular detection amplification and specific probes [14]. These last systems are usually not open and only allow detection of one or a few specific targets; however, they may provide no information about presumptive antibiotic susceptibility (i.e., detection of MRSA) [15], [16].…”

Section: Introductionmentioning

confidence: 99%

Direct Identification of Bacteria in Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption Ionisation Time-of-Flight Mass Spectrometry

2009

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BackgroundWith long delays observed between sampling and availability of results, the usefulness of blood cultures in the context of emergency infectious diseases has recently been questioned. Among methods that allow quicker bacterial identification from growing colonies, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry was demonstrated to accurately identify bacteria routinely isolated in a clinical biology laboratory. In order to speed up the identification process, in the present work we attempted bacterial identification directly from blood culture bottles detected positive by the automate.Methodology/Principal FindingsWe prospectively analysed routine MALDI-TOF identification of bacteria detected in blood culture by two different protocols involving successive centrifugations and then lysis by trifluoroacetic acid or formic acid. Of the 562 blood culture broths detected as positive by the automate and containing one bacterial species, 370 (66%) were correctly identified. Changing the protocol from trifluoroacetic acid to formic acid improved identification of Staphylococci, and overall correct identification increased from 59% to 76%. Lack of identification was observed mostly with viridans streptococci, and only one false positive was observed. In the 22 positive blood culture broths that contained two or more different species, only one of the species was identified in 18 samples, no species were identified in two samples and false species identifications were obtained in two cases. The positive predictive value of bacterial identification using this procedure was 99.2%.Conclusions/SignificanceMALDI-TOF MS is an efficient method for direct routine identification of bacterial isolates in blood culture, with the exception of polymicrobial samples and viridans streptococci. It may replace routine identification performed on colonies, provided improvement for the specificity of blood culture broths growing viridans streptococci is obtained in the near future.

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Fluorescence in situ hybridisation (FISH) accelerates identification of Gram-positive cocci in positive blood cultures

Cited by 58 publications

References 33 publications

Molecular-based Screening for Perinatal Group B Streptococcal Infection: Implications for Prevention and Therapy

Molecular-based Screening for Perinatal Group B Streptococcal Infection: Implications for Prevention and Therapy

Molecular microbiological methods in the diagnosis of neonatal sepsis

Direct Identification of Bacteria in Positive Blood Culture Bottles by Matrix-Assisted Laser Desorption Ionisation Time-of-Flight Mass Spectrometry

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