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Genetic Variability among Group A and B Respiratory Syncytial Virus Isolates from a Large Referral Hospital in New Delhi, India

机译:来自印度新德里一家大型转诊医院的A和B组呼吸道合胞病毒分离株之间的遗传变异性

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摘要

Respiratory syncytial virus (RSV) is an important childhood pathogen of acute lower respiratory infections in developed and developing countries. The molecular epidemiology of RSV in India is largely unknown. The present study was undertaken to standardize and evaluate reverse transcription-PCR (RT-PCR) for the rapid and simultaneous detection of RSV groups A and B in clinical samples and to study intragroup genetic variability. RT-PCR was evaluated by comparing the results of seminested RT-PCR with centrifugation-enhanced cultures on 200 nasopharyngeal aspirates from children with acute lower respiratory infections. RSV was isolated in 34 nasopharyngeal aspirates by centrifugation-enhanced cultures and identified in 45 samples by RT-PCR. In 15 samples RSV was identified by seminested RT-PCR alone and in four by centrifugation-enhanced cultures alone. Of the 45 samples positive for RSV by nested PCR, 15 belonged to group A, 29 to group B, and one sample suggested a mixed infection. Group B RSV predominated in both years of the 2-year study. Genetic variability within RSV groups was studied by restriction fragment analysis of 35 PCR products. Among both group A and group B RSV, two different composite patterns were observed. Thus, RSV was found to be a major pathogen of acute lower respiratory tract infections in India, as it was detected in 24.5% of children by RT-PCR. RT-PCR provides a sensitive method for detection and typing of RSV group A and B viruses in clinical samples as well as a means to study intragroup variations. However, a higher sensitivity of detection of RSV in clinical samples can be obtained by its combination with additional techniques, such as virus cultivation.
机译:在发达国家和发展中国家,呼吸道合胞病毒(RSV)是急性下呼吸道感染的重要儿童期病原体。在印度,RSV的分子流行病学尚不清楚。本研究旨在标准化和评估逆转录PCR(RT-PCR),用于快速和同时检测临床样品中的RSV组A和B,并研究组内遗传变异性。通过将半嵌套式RT-PCR结果与离心增强培养物对来自急性下呼吸道感染儿童的200例鼻咽抽吸物进行比较,评估了RT-PCR。通过离心分离培养,在34份鼻咽抽吸物中分离出RSV,并通过RT-PCR在45个样品中鉴定出RSV。在15个样本中,仅通过半巢式RT-PCR鉴定出RSV,在四个样本中,仅通过离心增强培养法鉴定出RSV。通过巢式PCR检测到的RSV阳性的45个样本中,有15个属于A组,有29个属于B组,其中一个样本表明存在混合感染。在2年研究的两年中,B组RSV占主导地位。通过对35种PCR产物进行限制性片段分析,研究了RSV组内的遗传变异性。在A组和B组RSV中,观察到两种不同的复合模式。因此,在印度,RSV是急性下呼吸道感染的主要病原体,通过RT-PCR在24.5%的儿童中检测到。 RT-PCR提供了一种检测和分型临床样品中RSV A和B组病毒的灵敏方法,也是研究组内变异的一种手段。但是,通过将其与其他技术(例如病毒培养)结合使用,可以获得更高的临床样品中RSV检测灵敏度。

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