农业生物技术学报2011，Vol.19Issue(2)：388-392,5.DOI:10.3969/j.issn.1674-7968.2011.02.028
新型鸭呼肠孤病毒RT-PCR方法的建立与应用
Establishment and Its Application of RT-PCR for Detection of Novel duck reovirus
摘要
Abstract
To develop a RT-PCR assay for the detection of novel duck reovirus (NDRV) and apply the developed assay to isolated strains and artificial infected samples, according to NDRV-NP03 S3 gene in the GenBank(GenBank:GQ888710), a pair of primers for amplying NDRV specifical fragment were designed and synthesized.RT-PCR technique detecting the RNA of NDRV was constructed.RT-PCR results showed that a 586 bp specifical fragment could be isolated only from the NDRV-NP03 strain RNA, and the sensitivity of RT-PCR reached to 2 pg NDRV-RNA.And the negative results were achieved from the other viruses, Muscovy duck reovirus(MDRV), Avian reovirus(ARV), Infectious bursal disease virus(IBDV), Muscovy duck parvovirus(MDPV), Goose parvovirus(GPV),Duck paramyxovirus(DPMV) and Duck hepatitis virus(DHV).The positive rate of RT-PCR method for detecting viral pathogens in 8 field NDRV isolates and 3 samples of the liver and spleen tissues of ducklings which were artificially infected by the strain of NDRV-NP03 was 100％.The results indicate that this RT-PCR method is sensitive and specific for detecting NDRV, and can be used in NDRV clinical diagnosis and epidemiology investigation.关键词
新型鸭呼肠孤病毒/RT-PCR/检测Key words
Novel duck reovirus/ RT-PCR/ Detection引用本文复制引用
王劭,陈少莺,陈仕龙,程晓霞,林锋强,朱小丽,江斌,李兆龙..新型鸭呼肠孤病毒RT-PCR方法的建立与应用[J].农业生物技术学报,2011,19(2):388-392,5.基金项目
本研究由福建省公益类科研院所专项基金(No.2010R1101003-5)和福建省农科院动物传染病防控创新团队项目(No.STIF-Y02)共同资助 （No.2010R1101003-5）
