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Real-time PCR TaqMan assay for rapid screening of bloodstream infection
Authors:Hye-young Wang  Sunghyun Kim  Hyunjung Kim  Jungho Kim  Yeun Kim  Soon-Deok Park  Hyunwoo Jin  Yeonim Choi  Young Uh  Hyeyoung Lee
Institution:1. M&D, Inc., Wonju Eco Environmental Technology Center, Wonju, Gangwon, 200-722, Republic of Korea
2. Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Gangwon, 220-710, Republic of Korea
3. Institute for Life Science and Biotechnology, Yonsei University, Seoul, 120-749, Republic of Korea
4. Department of Laboratory Medicine, Yonsei University Wonju College of Medicine, Wonju, Gangwon, 220-701, Republic of Korea
5. Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan, 609-757, Republic of Korea
6. Department of Biomedical Laboratory Science, Songho College, Hoengseong, 225-704, Republic of Korea
Abstract:

Background

Sepsis is one of the main causes of mortality and morbidity. The rapid detection of pathogens in blood of septic patients is essential for adequate antimicrobial therapy and better prognosis. This study aimed to accelerate the detection and discrimination of Gram-positive (GP) and Gram-negative (GN) bacteria and Candida species in blood culture samples by molecular methods.

Methods

The Real-GP®, -GN®, and -CAN® real-time PCR kit (M&D, Wonju, Republic of Korea) assays use the TaqMan probes for detecting pan-GP, pan-GN, and pan-Candida species, respectively. The diagnostic performances of the real-time PCR kits were evaluated with 115 clinical isolates, 256 positive and 200 negative blood culture bottle samples, and the data were compared to results obtained from conventional blood culture.

Results

Eighty-seven reference strains and 115 clinical isolates were correctly identified with specific probes corresponding to GP-bacteria, GN-bacteria and Candida, respectively. The overall sensitivity and specificity of the real-time PCR kit with blood culture samples were 99.6% and 89.5%, respectively.

Conclusions

The Real-GP®, -GN®, and -CAN® real-time PCR kits could be useful tools for the rapid and accurate screening of bloodstream infections (BSIs).
Keywords:
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