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首页> 外文期刊>Journal of Clinical Microbiology >Real-Time Reverse Transcription-PCR for Detection of Rotavirus and Adenovirus as Causative Agents of Acute Viral Gastroenteritis in Children
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Real-Time Reverse Transcription-PCR for Detection of Rotavirus and Adenovirus as Causative Agents of Acute Viral Gastroenteritis in Children

机译：实时逆转录-PCR检测轮状病毒和腺病毒作为儿童急性病毒性胃肠炎的病原体

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Viral pathogens are the most common cause of gastroenteritis in developed countries. Human rotavirus and adenovirus infections are major causes of acute outbreaks and sporadic cases of gastroenteritis, occurring primarily among children less than 2 years of age. Patient hospitalization is often required, with enormous infection control implications. This work describes the development of real-time PCR assays for the detection of group F adenovirus, rotavirus A, and rotavirus C from stool specimens. Two hundred twenty stool samples from pediatric patients exhibiting symptoms of diarrhea and/or vomiting were examined. PCR results were compared with those of virus detection by electron microscopy and latex agglutination antigen detection. The incorporation of an internal-control RNA that was spiked into individual stool extracts functioned as an internal validation for the reporting of PCR-negative results. Rotavirus C was not detected by real-time PCR in the patient stool samples examined. Real-time reverse transcription-PCR resulted in 175% and 111% increases in the rates of detection of adenovirus F and rotavirus A, respectively, compared with latex agglutination testing. Molecular detection increased the number of stool specimens in which causative agents of gastroenteritis were identified by 155% compared to electron microscopy. Genotyping of a proportion of the rotavirus and adenovirus strains identified only genotype G1 rotavirus and both adenovirus genotypes 40 and 41 in circulation within the patient cohort examined. The results highlight the significance of rapid molecular methods for the routine screening of stool samples in hospital laboratories to provide rapid definitive diagnoses.
机译：在发达国家，病毒病原体是胃肠炎的最常见原因。人类轮状病毒和腺病毒感染是急性暴发和肠胃炎偶发病例的主要原因，主要发生在2岁以下的儿童中。通常需要患者住院治疗，这对控制感染具有重大意义。这项工作描述了实时PCR检测方法的发展，用于从粪便标本中检测F组腺病毒，轮状病毒A和轮状病毒C。检查了来自小儿患者的220份粪便样本，这些样本表现出腹泻和/或呕吐症状。将PCR结果与通过电子显微镜和乳胶凝集抗原检测的病毒检测结果进行比较。掺入单个粪便提取物中的内部对照RNA的功能可作为内部验证，用于报告PCR阴性结果。实时PCR未在患者粪便样本中检测到轮状病毒C。与乳胶凝集试验相比，实时逆转录PCR分别使腺病毒F和轮状病毒A的检出率提高了175％和111％。与电子显微镜相比，分子检测使粪便标本的数量增加了155％，粪便标本中鉴定出肠胃炎的病原体。一部分轮状病毒和腺病毒株的基因分型仅确定了基因G1轮状病毒以及腺病毒基因型40和41在所研究的患者队列中的循环中。结果突出了快速分子方法对于医院实验室常规筛查粪便样本以提供快速确定性诊断的重要性。

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