摘要
目的建立仙台病毒(SV)RT-PCR检测方法,并对灰仓鼠仙台病毒感染情况进行调查。方法根据NCBI发表的SV(gi:9627219)基因组序列设计引物,建立RT-PCR方法,对方法的特异性和灵敏性进行验证,并用该方法检测60份灰仓鼠的肺脏样本。结果建立的SV RT-PCR方法显示有较好的敏感性和特异性:以仙台病毒为模板扩增产生197 bp的单一目的条带,经测序比对与NCBI数据库中SV相关序列的一致率为98%,而以猴副流感病毒(SV5)、犬瘟热病毒、小鼠肺炎病毒、呼肠孤病毒III型及腮腺炎病毒为对照无任何条带产生;能检出的SVcDNA最低浓度是96.8 ng/mL;用该方法检测60份灰仓鼠,SV的感染率为3.33%(2/60)。结论建立的SV RT-PCR方法可用于实验类啮齿动物动物SV的常规检测,自然条件下灰仓鼠感染SV的问题不容忽视。
Objective To establish a RT-PCR detection method for Sendai virus and to investigate the infection status in Cricetulus migratorius.Methods To design PCR primers according to the Sendai virus（gi：9627219）genome in NCBI and establish a RT-PCR detection method using the Sendai virus strain from our department.To screen the Sendai virus infection in sixty Cricetulus migratorius lung samples by the established RT-PCR procedure.Results The RT-PCR method established was more sensitive and specific： there was a 197 bp single band when SV was used as template and no any band occurred when SV5,CDV,PVM,Reo3 and MuV were used as templates.Sequencing results revealed that the 197 bp nucleotides showed 98% coincidence with the SV sequence published by NCBI.The assay could detect as low as 96.8 ng/mL SV cDNA.Results showed that the infection rate of Sendai virus was 3.33%（2/60）in Cricetulus migratorius.Conclusions The established RT-PCR detection technique is sensitive,specific and rapid,and can be used to detect Sendai virus routinely in laboratory rodent animals.The positive results of Sendai virus infection show that Sendai virus infection in Cricetulus migratorius can not be ignored.
出处
《中国比较医学杂志》
CAS
2012年第12期18-22,共5页
Chinese Journal of Comparative Medicine
基金
国家科技支撑计划"野生动物人工种群的生物净化及相关疾病动物模型的建立与评价"(2009BAI83B02)
关键词
仙台病毒
感染
检测
逆转录-聚合酶链反应
灰仓鼠
Sendai virus
Infection
Detection
Reverse transcriptase polymerase chain reaction
Cricetulus migratorius
