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目的本文主要研究建立一种酶切图谱法从日本历年本土C1、D3、D5基因型麻疹病毒中鉴别中国输入株H1基因型麻疹病毒,即逆转录多聚酶链式反应—限制性片段长度多态性分析方法(RTPCRRFLP)。方法对C1、D3、D5及H1基因型麻疹病毒H基因全长的序列进行分析,寻找能够从日本本土株中鉴别出输入株H1基因型麻疹病毒的限制性内切酶酶切位点,建立RTPCRRFLP方法。同时,我们又应用多株已经过序列分析确定为C1、D3、D5、H1基因型的麻疹病毒用该RTPCRRFLP方法进行验证。结果应用该RTPCRRFLP方法所得结果与序列分析方法结果一致。结论该RTPCRRFLP方法是一种简单、快速又实用的从日本本土株中筛查H1基因型输入株的方法。
AIM: To establish a restriction map to identify H1 genotypes of imported Chinese isolates from C1, D3 and D5 genotypes of measles viruses in Japan over the calendar year, namely, reverse transcriptase polymerase chain reaction-restriction fragment length polymorphism Analysis Method (RTPCRRFLP). Methods The sequences of the full-length H gene of C1, D3, D5 and H1 genotypes were analyzed to find out the restriction enzyme sites that could identify the imported H1 strain measles virus from Japanese indigenous strains. RTPCRRFLP method. In the meantime, we used multiple strains of measles virus that have been identified as C1, D3, D5, and H1 genotypes by sequence analysis to verify the RTPCRRFLP method. Results The results obtained by using this RTPCRRFLP method were consistent with the results of sequence analysis. Conclusion The RTPCRRFLP method is a simple, rapid and practical method for screening H1 genotype imported strains from Japanese indigenous strains.