Open AccessRapid diagnosis of sepsis with TaqMan‐Based multiplex real‐time PCR
Author(s) -
Liu ChangFeng,
Shi XinPing,
Chen Yun,
Jin Ye,
Zhang Bing
Publication year - 2018
Publication title -
journal of clinical laboratory analysis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 50
eISSN - 1098-2825
pISSN - 0887-8013
DOI - 10.1002/jcla.22256
Subject(s) - taqman , multiplex , sepsis , real time polymerase chain reaction , multiplex polymerase chain reaction , blood culture , bacteria , biology , microbiology and biotechnology , medicine , polymerase chain reaction , immunology , gene , bioinformatics , antibiotics , genetics
Background The survival rate of septic patients mainly depends on a rapid and reliable diagnosis. A rapid, broad range, specific and sensitive quantitative diagnostic test is the urgent need. Thus, we developed a TaqMan‐Based Multiplex real‐time PCR assays to identify bloodstream pathogens within a few hours. Methods Primers and TaqMan probes were designed to be complementary to conserved regions in the 16S rDNA gene of different kinds of bacteria. To evaluate accurately, sensitively, and specifically, the known bacteria samples (Standard strains, whole blood samples) are determined by TaqMan‐Based Multiplex real‐time PCR . In addition, 30 blood samples taken from patients with clinical symptoms of sepsis were tested by TaqMan‐Based Multiplex real‐time PCR and blood culture. Results The mean frequency of positive for Multiplex real‐time PCR was 96% at a concentration of 100 CFU /mL, and it was 100% at a concentration greater than 1000 CFU /mL. All the known blood samples and Standard strains were detected positively by TaqMan‐Based Multiplex PCR , no PCR products were detected when DNA s from other bacterium were used in the multiplex assay. Among the 30 patients with clinical symptoms of sepsis, 18 patients were confirmed positive by Multiplex real‐time PCR and seven patients were confirmed positive by blood culture. Conclusion TaqMan‐Based Multiplex real‐time PCR assay with highly sensitivity, specificity and broad detection range, is a rapid and accurate method in the detection of bacterial pathogens of sepsis and should have a promising usage in the diagnosis of sepsis.