Japanese Journal of Infectious Diseases
Online ISSN : 1884-2836
Print ISSN : 1344-6304
Detection of Uncommon Enteric Bacterial Pathogens from Acute Diarrheal Specimens using SYBR-Green Real Time PCR
Goutam ChowdhuryOscar C. StineKrishnan RajendranAsish K. MukhopadhyayKeinosuke OkamotoThandavarayan Ramamurthy
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JOURNALS FREE ACCESS Advance online publication

Article ID: JJID.2018.142

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Abstract

Acute diarrheal disease is a major health problem and second most common cause of death in children under five years of age. Conventional diagnostic methods are laborious, time consuming and occasionally inaccurate. We examined SYBR-Green real-time PCR for the detection of ten uncommon bacterial pathogens using fecal specimens from acute diarrheal patients. In SYBR-Green real-time PCR, the products formed were identified based on melting point temperature (Tm) curve analysis and was validated with the respective reference strain. In a retrospective study, 1184 tested stool specimens by conventional culture methods were included. Enterotoxigenic Bacteriodes fragilis was detected more (6.7%) followed by enterotoxigenic Bacillus cereus (5.1%), Clostridium perfringens (3.9%) and Aeromonas hydrophila (3.8%). In the prospective study, A. hydrophila, S. aureus, and C. perfringens, were predominantly detected in >5 years age group by the real-time PCR. The real-time PCR assay is comprehensive, rapid, accurate and well suited for surveillance or diagnostic purpose for detection of uncommon bacterial pathogens and also helpful in initiating appropriate care and thereby reducing the risk of the patients.

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