Tatte Vaishali S, Gopalkrishna Varanasi
Enteric Viruses Group, National Institute of Virology, Pune, India.
Access Microbiol. 2019 Mar 29;1(2):e000010. doi: 10.1099/acmi.0.000010. eCollection 2019.
Enteric viruses play a major role in causing diarrhea in children. Early identification of the causative pathogen is still a challenge in the clinical laboratory. A multiplex PCR assay is a useful tool to screen a large number of clinical samples especially in an outbreak situation. In this study, a multiplex reverse transcription (RT)-PCR assay was developed to detect nine enteric viruses such as group A rotavirus, norovirus GGII, sapovirus, adenovirus, astrovirus, aichivirus, parechovirus, bocavirus and enterovirus in clinical samples of diarrheal cases. Stool samples (=185) collected from infants and children with acute gastroenteritis cases in Pune, western India were analysed for nine different enteric viruses by currently developed multiplex RT- PCR. Predominance of group A rotavirus (76%) followed by enterovirus (11.5%), astrovirus (4.5%), adenovirus (2.7%) and norovirus GII (1.6%) was observed. A total of 44.8 % (82/185) samples analysed by this method showed high frequency of mixed infections. These results highlighted high prevalence and diversity of different enteric viruses in children. The multiplex PCR showed good concordance with monoplex RT-PCR for detection of these enteric viruses in clinical samples. This is the first report on the development of a multiplex RT-PCR assay for detection of multiple enteric viruses in diarrheal diseases from India.
肠道病毒在引发儿童腹泻方面起着主要作用。在临床实验室中，早期识别致病病原体仍是一项挑战。多重PCR检测是筛选大量临床样本的有用工具，尤其是在疫情暴发的情况下。在本研究中，开发了一种多重逆转录（RT）-PCR检测方法，用于检测腹泻病例临床样本中的九种肠道病毒，如A组轮状病毒、诺如病毒GGII、札幌病毒、腺病毒、星状病毒、艾柯病毒、帕利病毒、博卡病毒和肠道病毒。从印度西部浦那的急性肠胃炎婴幼儿和儿童中收集了185份粪便样本，通过目前开发的多重RT-PCR对九种不同的肠道病毒进行分析。观察到A组轮状病毒占主导地位（76%），其次是肠道病毒（11.5%）、星状病毒（4.5%）、腺病毒（2.7%）和诺如病毒GII（1.6%）。通过该方法分析的样本中，共有44.8%（82/185）显示出高频率的混合感染。这些结果突出了儿童中不同肠道病毒的高流行率和多样性。多重PCR在临床样本中检测这些肠道病毒时与单重RT-PCR显示出良好的一致性。这是印度关于开发用于检测腹泻疾病中多种肠道病毒的多重RT-PCR检测方法的首份报告。
