Genetic variability among Group A and B respiratory syncytial virus isolates from a large referral hospital in New Delhi, India.
Genetic variability among Group A and B respiratory syncytial virus isolates from a large referral hospital in New Delhi, India.
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印度新德里一家大型转诊医院分离出的 A 组和 B 组呼吸道合胞病毒的遗传变异。
DOI：
10.1128/jcm.41.6.2311-2316.2003
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发表时间：
2003
影响因子：
9.4
通讯作者：
Broor,Shobha
中科院分区：
文献类型：
--
作者：
Rajala,MaitreyiS;Sullender,WayneM;Prasad,AK;Dar,Lalit;Broor,Shobha
呼吸道合胞病毒(RSV)是发达国家和发展中国家儿童急性下呼吸道感染的重要病原体。在印度，RSV的分子流行病学在很大程度上是未知的。本研究旨在规范和评价逆转录聚合酶链式反应(RT-PCR)快速同时检测临床标本中RSV A、B组的方法，并研究其组内遗传变异。通过比较200例急性下呼吸道感染患儿鼻咽抽吸物的半套式RT-PCR和离心增强培养的结果，对RT-PCR进行评价。34份鼻咽抽吸物经离心强化培养分离出呼吸道合胞病毒，用RT-PCR方法鉴定出45份样本。在15个样本中，仅通过半套式RT-PCR检测到RSV，在4个样本中仅通过离心力增强培养检测到RSV。45份套式聚合酶链式反应阳性标本中，15份为A组，29份为B组，1份为混合感染。在两年的研究中，B组RSV占主导地位。通过对35个聚合酶链式反应产物的限制性片段分析，研究了RSV组内的遗传变异。在A组和B组RSV之间，观察到两种不同的复合模式。因此，呼吸道合胞病毒被发现是印度急性下呼吸道感染的主要病原体，通过RT-PCR在24.5%的儿童中检测到该病毒。RT-PCR为临床标本中呼吸道合胞病毒A组和B组病毒的检测和分型提供了一种敏感的方法，也是研究组内变异的一种手段。然而，通过与其他技术相结合，如病毒培养，可以获得更高的临床样本中RSV的检测灵敏度。
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.