摘要
目的建立新城疫病毒(Newcastle disease virus,NDV)一步法RT-PCR检测方法,并进行验证及初步应用。方法根据NDV F基因序列,设计合成了1对特异性引物,建立一步法RT-PCR检测方法,并验证其特异性及敏感性;应用建立的方法对23份疑似新城疫(Newcastle disease,ND)病料进行检测,并与血凝及血凝抑制试验检测结果进行比对。结果建立的一步法RT-PCR同时检测NDV、传染性法氏囊病病毒(Infectious bursal disease virus,IBDV)和禽流感病毒(Avian influenza virus,AIV)H9亚型,仅NDV为阳性,IBDV和AIV H9均为阴性;该方法最低可检出约4 pg的NDV RNA;23份疑似ND病料,一步法RT-PCR检出11份阳性,血凝及血凝抑制试验检出13份阳性,两种方法的阳性符合率为85%。结论建立了NDV一步法RT-PCR检测方法,该方法特异性良好,敏感度较高,用时较短,可从分子水平上对NDV进行早期快速诊断和流行病学调查。
Objective To develop a one-step RT-PCR assay for Newcastle disease virus （NDV）. Methods A pair of specific primers was designed according to the sequences of F gene of NDV, based on which a one step RT-PCR assay was developed and verified for specificity and sensitivity. Twenty-three clinic suspected samples were detected by the developed RT-PCR method, of which the results were compared with those by hemagglutination （HA） and hemagglutination inhibition （HI） tests. Results By developed RT-PCR method, the detection result of NDV was positive, while those of infectious bursal disease virus （IBDV） and avian influenza virus （AIV） subtype H9 were negative. The minimum detection limit of the method was 4 pg NDV RNA. Of the 23 clinic suspected samples, 11 were judged as positive by RT-PCR, and 13 by HA and HI tests, indicating a positive coincidence rate of 85%. Conclusion One-step RT-PCR assay for NDV was developed, which was specific, sensitive and time-saving, and might be used for early rapid diagnosis and epidemiological investigation of NDV at molecular level.
出处
《中国生物制品学杂志》
CAS
CSCD
2013年第2期265-267,共3页
Chinese Journal of Biologicals
关键词
新城疫病毒
逆转录聚合酶链反应
Newcastle disease virus （NDV）
Reverse transcription-polymerase chain reaction （RT-PCR）
