呼吸道合胞病毒PCR序列分析方法的建立和应用

The establishment and application of the method to respiratory syncytial virus polymerase chain reaction sequencing strategy

摘要: 目的：探讨呼吸道合胞病毒cDNA（RSVcDNA）序列分析的方法。方法：将PCR扩增与核酸序列分析技术相结合，以γ³²P-ATP标记T₇Promoter为测序引物，Taq DNA聚合酶直接测序。结果：测序梯清晰，可读性好。RSV NS和N基因区虽较稳定，但仍有颠换突变。结论：RSV的变异性决定了它对人类的反复感染，PCR测序方法较简便，在临床标本的检测和变异的观察中有重要意义。

Abstract: Objective: To search the method of respiratory syncytial virus cDNA（RSV cDNA） sequencing strategy. Methods: The polymerase chain reaction （PCR） was combined with the sequencing strategy by using γ³²P-ATP labeled T 7 Promoter as sequencing primer and the thermostable DNA polymerose as directed sequencing strategy.Results: The sequencing ladder was clear and readable. Although NS、N genes of RSV were rather stable, there were still exchanging mutations.Conclusion: The PCR sequencing strategy is a simple and reliable method for detecting RSV and its variability in patients with recurrent RSV infections.