Comparison of nested, multiplex, qPCR; FISH; SeptiFast and blood culture methods in detection and identification of bacteria and fungi in blood of patients with sepsisReport as inadecuate




Comparison of nested, multiplex, qPCR; FISH; SeptiFast and blood culture methods in detection and identification of bacteria and fungi in blood of patients with sepsis - Download this document for free, or read online. Document in PDF available to download.

BMC Microbiology

, 14:313

Clinical microbiology and vaccines

Abstract

BackgroundMicrobiological diagnosis of sepsis relies primarily on blood culture data. This study compares four diagnostic methods, i.e. those developed by us: nested, multiplex, qPCR qPCR and FISH with commercial methods: SeptiFast Roche SF and BacT-ALERT® 3D blood culture system bioMérieux. Blood samples were derived from adult patients with clinical symptoms of sepsis, according to SIRS criteria, hospitalized in the Intensive Care Unit.

ResultsUsing qPCR, FISH, SF, and culture, microbial presence was found in 71.8%, 29.6%, 25.3%, and 36.6% of samples, respectively. It was demonstrated that qPCR was significantly more likely to detect microorganisms than the remaining methods; qPCR confirmed the results obtained with the SF kit in all cases wherein bacteria were detected with simultaneous confirmation of Gram-typing. All data collected through the FISH method were corroborated by qPCR.

ConclusionsThe qPCR and FISH methods described in this study may constitute alternatives to blood culture and to the few existing commercial molecular assays since they enable the detection of the majority of microbial species, and the qPCR method allows their identification in a higher number of samples than the SF test. FISH made it possible to show the presence of microbes in a blood sample even before its culture.

KeywordsNested Multiplex qPCR FISH SeptiFast Blood culture Sepsis Electronic supplementary materialThe online version of this article doi:10.1186-s12866-014-0313-4 contains supplementary material, which is available to authorized users.

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Author: Tomasz Gosiewski - Agnieszka Flis - Agnieszka Sroka - Anna Kędzierska - Agata Pietrzyk - Jolanta Kędzierska - Rafał Drwi

Source: https://link.springer.com/







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