Detection Of Uncommon Enteric Bacterial Pathogens From Human Diarrheal Specimens By SYBR-Green Real Time PCR | 51368
Journal of Infectious Diseases & Therapy
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Acute diarrheal disease is still a major health problem and second most common cause of death worldwide in children under five
years of age. Most of the morbidity occurs in low-income countries, where the etiologies and epidemiology of gastroenteritis
remain incompletely understood. Diarrhea can be caused by a range of pathogens, including several bacteria. Conventional diagnostic
methods, such as culture, biochemical tests and enzyme-linked immunosorbent assay (ELISA) are laborious and time consuming.
We used SYBR-Green real time PCR assay targeting 10 uncommon diarrheagenic bacterial pathogens (S. aureus, Enterotoxigenic B.
cereus, C. perfringens, C. difficile, L. monocytogenes, P. shigelloides, Y. enterocolitica, Enterotoxigenic B. fragilis, A. hydrophila and P.
alcalifaciens) directly in fecal specimens from patients admitted infectious diseases hospital with acute diarrhea in Kolkata, India. The
products formed were identified based on melting point temperature (Tm) curve analysis. The assay was first validated with reference
strains or isolates and exhibited a limit of detection of 103 to 105 CFU/gm of stool for each pathogen. A total of 1184 clinical fecal
specimens from individual with diarrhea, previously cultured for enteric pathogens were evaluated. Enterotoxigenic B. fragilis was
detected highest number about 80 (6.75%) followed by enterotoxigenic B. cereus 60 (5.06%), C. perfringens 46 (3.88%), A. hydrophila
45 (3.80%), P. alcalifaciens 44 (3.71%), P. shigelloides 39 (3.29%), C. difficile 39 (3.29%), L. monocytogenes 38 (3.20%), S. aureus 23
(1.94%) and Y. enterocolitica 14 (1.8%) respectively. We found SYBR-Green real time PCR assay for simultaneous detection of 10
target pathogens to be comprehensive, rapid, inexpensive and accurate, of high selectivity and is well suited for surveillance or clinical