关键词: 猴副流感病毒(Simian parainfluenza virus 5, SV5), 核酸检测, RT-PCR

Abstract: To detect the presence of Simian parainfluenza virus 5(SV5) in laboratory animals and related biological products or materials,a pair of primers that amplify the SH gene were designed and synthesized.After the best reaction conditions were tested out,RT-PCR technique was established for the detection of RNA of SV5.By nucleic acid sequence analysis,sequence homology of RT-PCR positive fragment and the reported SV5 WR-21005 strain achieved 97%,which testified that the method was specific for the detection of SV5 nucleic acid sequence.Sensitivity of the method was determined as the minimum RNA concentration of 2.1 ng/μL and the minimum virus titer of 3.98CCID_(50)/0.1 mL.For initial application of the method,samples including organs of simian and hamster's,biological products from simian(OPV) and five commonly used passage cells from simian kidney were tested by the RT-PCR method for the SV5 nucleotide gene.Results came out that no SV5 nucleotide gene was detected in above specimens.The results showed that the SV5 RT-PCR technique we established provided a diagnosis method of SV5,and also a technique for SV5 molecular epidemiologic survey.

Key words: Simian parainfluenza virus 5, Nucleic acid detecton, RT-PCR