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A Comparative Study for the Efficient Detection of Norovirus from Drinking Water by RT-PCR and Real-Time PCR

机译:RT-PCR和实时荧光定量PCR检测饮用水中诺如病毒的比较研究

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Noroviruses (NoV), of the family Caliciviridae, are the most important cause of outbreaks of acute gastroenteritis worldwide. Water can be a source of disease outbreaks. Contamination of drinking water takes place almost exclusively through sewage that contains enteric pathogens. In India, no attempt has been made to detect the presence of norovirus in drinking water earlier and the present study is the first attempt in Chennai to identify norovirus from drinking water. In this study, 100 samples were collected and were concentrated for norovirus using granular activated carbon. The main objective of the study is to compare the efficiency of reverse transcriptase-polymerase chain reaction (RT-PCR) with that of real-time polymerase chain reaction. Conventional RT-PCR was performed using the primers NI and E3 and the results were confirmed by agarose gel electrophoresis as well as by slot blot analysis. Real-time PCR assay was performed using SYBR green RT-PCR Kit. Out of the 100 samples, 18% of the samples were positive by conventional RT-PCR, while 20% were positive by real-time PCR reactions. The real-time PCR assays provide rapid, sensitive, and reliable detection of norovirus and proved to be useful for routine monitoring of drinking water samples.
机译:杯状病毒科的诺如病毒(NoV)是全世界急性胃肠炎暴发的最重要原因。水可能是疾病暴发的源头。饮用水的污染几乎完全是通过含有肠道病原体的污水进行的。在印度,尚未尝试过更早地检测饮用水中是否存在诺如病毒,而本研究是金奈首次从饮用水中鉴定诺如病毒的尝试。在这项研究中,收集了100个样品,并使用颗粒状活性炭对诺如病毒进行了浓缩。该研究的主要目的是比较逆转录聚合酶链反应(RT-PCR)和实时聚合酶链反应的效率。使用引物NI和E3进行常规RT-PCR,结果通过琼脂糖凝胶电泳和狭缝印迹分析得到证实。使用SYBR green RT-PCR Kit进行实时PCR分析。在100个样本中,常规RT-PCR阳性的样本占18%,而实时PCR反应阳性的样本占20%。实时PCR检测可快速,灵敏和可靠地检测诺如病毒,并被证明可用于常规监测饮用水样品。

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