仙台病毒核酸测序检测方法的建立

目的 建立仙台病毒(SeV)的核酸测序检测方法.方法 根据SeV序列设计覆盖不同毒株的通用引物,然后优化成测序引物并摸索建库条件,进行核酸测序和结果分析.结果 获得1对特异性强的通用引物,序列为:上游引物5’-GCTGCAAAACGCTGTGGG-3’,下游引物5’-TGGRACYTCAGAAAGAATRGG-3’;建库条件优化为:第一轮以cDNA为模板用通用引物扩增,第二轮以第一轮产物为模板用测序引物扩增.测序分析可以有效地将SeV与其他微生物区分开,并且精确到TianJin亚株.结论 建立了SeV核酸测序检测方法,为后续SeV感染实验动物的检验检疫提供依据.

Development of Detection Method for Sendai Virus by Nucleic Acid Sequencing

Objective To develop the method for detection of Sendai virus by nucleic acid sequencing.Methods General primers according to Sendai virus genome sequences were designed and optimized to sequencing primers.Then the best condition of creating library was established.This method was verified by nucleic acid sequencing and analysis.Results A pair of highly specific general primers was designed.The sequences are respectively 5'-GCTGCAAAACGCTGTGGG-3'and 5'-TGGRACYT-CAGAAAGAATRGG-3',and the sequencing primers were established by ligated adaptor sequences to the 5'-end of general primers.The established condition of creating library and library products were obtained by the first round amplification using general primers and the second round using sequencing primers.Sendai virus was effectively distinguish from other organisms by nucleic acid sequencing and analysis,and was precised to TianJin strain.Conclusion A detection method was developed for Sendai virus by nucleic acid sequencing,which may be as the base for microbiological detection in laboratory animals.