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Detection of Clostridium difficile toxin A/B genes by multiplex real-time PCR for the diagnosis of C. difficile infection

Authors
 Heejung Kim  ;  Seok Hoon Jeong  ;  Myungsook Kim  ;  Yangsoon Lee  ;  Kyungwon Lee 
Citation
 JOURNAL OF MEDICAL MICROBIOLOGY, Vol.61(2) : 274-277, 2012 
Journal Title
 JOURNAL OF MEDICAL MICROBIOLOGY 
ISSN
 0022-2615 
Issue Date
2012
MeSH
Bacterial Proteins/analysis* ; Bacterial Proteins/genetics ; Bacterial Toxins/analysis* ; Bacterial Toxins/genetics ; Bacteriological Techniques/methods ; Clostridium Infections/diagnosis* ; Clostridium Infections/microbiology ; Clostridium difficile/genetics ; Clostridium difficile/isolation & purification* ; Enterotoxins/analysis* ; Enterotoxins/genetics ; Feces/microbiology ; Humans ; Molecular Diagnostic Techniques/methods* ; Multiplex Polymerase Chain Reaction/methods* ; Predictive Value of Tests ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction/methods* ; Sensitivity and Specificity
Keywords
Bacterial Proteins/analysis* ; Bacterial Proteins/genetics ; Bacterial Toxins/analysis* ; Bacterial Toxins/genetics ; Bacteriological Techniques/methods ; Clostridium Infections/diagnosis* ; Clostridium Infections/microbiology ; Clostridium difficile/genetics ; Clostridium difficile/isolation & purification* ; Enterotoxins/analysis* ; Enterotoxins/genetics ; Feces/microbiology ; Humans ; Molecular Diagnostic Techniques/methods* ; Multiplex Polymerase Chain Reaction/methods* ; Predictive Value of Tests ; Reagent Kits, Diagnostic ; Real-Time Polymerase Chain Reaction/methods* ; Sensitivity and Specificity
Abstract
Toxigenic Clostridium difficile culture is considered to be the standard diagnostic method for the detection of C. difficile infection (CDI). Culture methods are time-consuming and although enzyme immunoassay is rapid and easy to use, it has low sensitivity. In the present study, the AdvanSure CD real-time (RT)-PCR kit (LG Life Sciences) was evaluated for its ability to detect C. difficile toxin A (tcdA) and B (tcdB) genes, simultaneously. A total of 127 fresh diarrhoeal stool specimens, submitted to the clinical microbiology laboratory for C. difficile culture, were tested. C. difficile toxins and toxin genes were detected with a VIDAS C. difficile A&B (VIDAS-CDAB) enzyme-linked fluorescent immunoassay (ELFA) and the AdvanSure RT-PCR kit, respectively, according to the manufacturers’ instructions. Their performance was compared with a standard toxigenic culture method as a reference. The sensitivity, specificity and positive and negative predictive values using the AdvanSure RT-PCR kit were 100 %, 98.3 %, 84.6 % and 100 %, respectively, while those of the VIDAS-CDAB system were 63.6 %, 100 %, 100 % and 96.6 %, respectively. Four tcdA+/tcdB+ strains of C. difficile were detected with the AdvanSure RT-PCR kit, which offers comparable sensitivity and specificity to the reference method with a turnaround time of ~3 hours.
Full Text
http://jmm.sgmjournals.org/content/61/2/274.short
DOI
21959205
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Laboratory Medicine (진단검사의학교실) > 1. Journal Papers
Yonsei Authors
Kim, Heejung(김희정) ORCID logo https://orcid.org/0000-0002-0190-703X
Lee, Kyungwon(이경원) ORCID logo https://orcid.org/0000-0003-3788-2134
Lee, Yang Soon(이양순)
Jeong, Seok Hoon(정석훈) ORCID logo https://orcid.org/0000-0001-9290-897X
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/90961
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