Singh Taru, Das Shukla, Ramachandran V G, Dar Sajad Ahmad, Snehaa K, Saha Rumpa, Shah Dheeraj
Department of Microbiology, UCMS, Guru Teg Bahadur Hospital, New Delhi, India.
Department of Paediatrics, UCMS, Guru Teg Bahadur Hospital, New Delhi, India.
Indian J Med Microbiol. 2017 Apr-Jun;35(2):204-210. doi: 10.4103/ijmm.IJMM_16_21.
Diarrhoeagenic Escherichia coli (DEC) is associated with early death of children in developing countries and are being identified now as an important evolving pathogen. The objective of this study was to perform multiplex polymerase chain reaction (PCR) for simultaneous detection of six categories of DEC in two sets of PCR reactions using 11 virulent genes.
During 1-year study period, forty isolates each from outpatient, inpatient and healthy groups were collected from children. E. coli was identified using conventional biochemical methods. DNA extraction was done using kit, and the extracted DNA was used as a template for multiplex PCR.
Virulent genes of DEC were detected in 106 (88.33%) samples. Overall, elt and est were detected in 8.33% and 30.83% of specimens; typical, atypical enteropathogenic E. coli and bfp were detected in 13.33%, 29.16% and 19.16% specimens; eagg was detected in 39.16% and east in 13.33% specimens and stx and hyla were isolated in 1.66% specimens each. While diffusely adherent E. coli and enteroinvasive E. coli genes were not isolated.
Multiplex PCR is a rapid method for the simultaneous detection of 11 virulent genes of DEC at a time and it will provide a platform in understanding the diarrheal diseases in a more improved manner.
致泻性大肠杆菌(DEC)与发展中国家儿童的早期死亡有关,目前被确定为一种重要的不断演变的病原体。本研究的目的是进行多重聚合酶链反应(PCR),以在两组PCR反应中使用11个毒力基因同时检测六类DEC。
在为期1年的研究期间,从儿童中分别收集了门诊、住院和健康组的40株分离株。使用传统生化方法鉴定大肠杆菌。使用试剂盒进行DNA提取,并将提取的DNA用作多重PCR的模板。
在106份(88.33%)样本中检测到DEC的毒力基因。总体而言,elt和est分别在8.33%和30.83%的标本中被检测到;典型、非典型肠致病性大肠杆菌和bfp分别在13.33%、29.16%和19.16%的标本中被检测到;eagg在39.16%的标本中被检测到,east在13.33%的标本中被检测到,stx和hyla分别在1.66%的标本中被分离到。而弥漫性黏附性大肠杆菌和肠侵袭性大肠杆菌基因未被分离到。
多重PCR是一种可同时快速检测DEC的11个毒力基因的方法,它将为以更好的方式理解腹泻疾病提供一个平台。