Multiplex PCR for typing and subtyping influenza and respiratory syncytial viruses
Multiplex PCR for typing and subtyping influenza and respiratory syncytial viruses
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DOI：
10.1128/jcm.36.10.2990-2995.1998
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发表时间：
1998-10-01
影响因子：
9.4
通讯作者：
Zambon, MC
中科院分区：
文献类型：
--
作者：
Stockton, J;Ellis, JS;Zambon, MC
已开发的多重逆转录（RT）-PCR方法能够检测和分型甲型流感（H1 N1和H3 N2）和B病毒以及呼吸道合胞病毒（RSV）A型和B型，这些呼吸道临床样本是英格兰和威尔士流感样疾病国家社区监测计划的一部分。每种不同病原体的检测依赖于用多个引物组进行RT-PCR后在琼脂糖凝胶上区分5种不同大小的扩增产物。多重RT-PCR检测了65个鼻咽部的蜜蜂，从RSV分离和237个合并的鼻和咽喉拭子，流感病毒A（H1N1和H3 N2）或B病毒已通过病毒分离检测，以及40个呼吸道样本，其他病毒，包括巨细胞病毒或单纯疱疹病毒，肠道病毒，副流感病毒已生长。对于甲型和B型流感病毒以及A型和B型RSV的分型和亚型，多重RT-PCR与用特异性抗血清进行的常规分型和亚型分型结果具有极好的相关性（100%）。多重RT-PCR还可用于准确检测同一反应混合物中的一种以上病毒模板，从而可以用同一呼吸道标本鉴定病毒合并感染。
A multiplex reverse transcription (RT)-PCR method that has been developed is capable of detecting and subtyping influenza A (H1N1 and H3N2) and B viruses as well as respiratory syncytial virus (RSV) types A and B in respiratory clinical samples taken as part of a national community-based surveillance program of influenza-like illness in England and Wales. The detection of each different pathogen depended on distinguishing five amplification products of different sizes on agarose gels following RT-PCR with multiple primer sets. The multiplex RT-PCR was tested with 65 nasopharyngeal apirates from which RSV had been isolated and 237 combined nose and throat swabs from which influenza A (H1N1 and H3N2) or B virus had been detected by virus isolation, as well as 40 respiratory samples from which other viruses including cytomegalovirus lr herpes simplex virus, enteroviruses, and parainfluenza viruses had been grown. For the typing and subtyping of influenza A and B viruses and RSV types A and B, the multiplex RT-PCR gave an excellent (100%) correlation with the results of conventional typing and subtyping with specific antisera. Multiplex RT-PCR can also be used to accurately detect more than one viral template in the same reaction mixture, allowing viral coinfections to be identified with the same respiratory specimen.