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

BMC Microbiology

Methodology Article

Małgorzata Bulanda¹  Agnieszka Sroka²  Tomasz Gosiewski²  Agata Pietrzyk²  Anna Kędzierska³  Jolanta Kędzierska³  Agnieszka Flis⁴  Rafał Drwiła⁵ 

[1] Department Epidemiology of Infection, Chair of Microbiology Jagiellonian University Medical College, Czysta 18 Str., 31-121, Krakow, Poland;Department of Bacteriology, Microbial Ecology and Parasitology, Chair of Microbiology Jagiellonian University Medical College, Czysta 18 Str., 31-121, Krakow, Poland;Department of Microbiology of the University Hospital in Krakow, Kopernika 19 Str., 31-501, Krakow, Poland;Intensive Care Unit of the Cardiac Surgery Department, The John Paul II Hospital in Krakow, Pradnicka 80 Str., 31-202, Krakow, Poland;Intensive Care Unit of the Cardiac Surgery Department, The John Paul II Hospital in Krakow, Pradnicka 80 Str., 31-202, Krakow, Poland;Department of Anesthesiology and Intensive Care, Jagiellonian University, Kopernika 17 Str., 31-501, Medical College, Krakow, Poland;

关键词: Nested;    Multiplex qPCR;    FISH;    SeptiFast;    Blood culture;    Sepsis;   

DOI  :  10.1186/s12866-014-0313-4

 received in 2014-09-01, accepted in 2014-11-26,  发布年份 2014

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY   
© Gosiewski et al.; licensee BioMed Central. 2014