摘要
根据已发表的CSFV序列合成1对引物，以猪瘟病毒（CSFV）兔化弱毒株为模板，建立并优化了检测CSFV的RT—PCR方法，并与直接荧光抗体试验（DFA）进行比较，检测了15份临床疑似病料。结果如下：应用RT—PCR对CSFV兔化弱毒株RNA进行扩增，获得与预期大小相符长为509bp的特异性目的片段，敏感性达到10Pg的CSFV—RNA，对猪繁殖与呼吸综合征病毒（PRRSV）、猪细小病毒（PPV）、猪伪狂犬病毒（PRV）和猪2型圆环病毒（PCV2）进行同条件扩增均为阴性，RT—PCR产物测序结果与文献报道的CSFV不同毒株的核苷酸序列同源性达到95％～99％；15份临床疑似病料应用RT—PCR的检出率为66．7％，而应用免疫荧光试验的检出率为60．0％，二者的总符合率为80％。表明RT—PCR方法比DFA更敏感，两种方法均可用于猪瘟的快速诊断。
A pair of primers for RT -PCR to detect the classical swine fever virus （CSFV） were synthesized according to the published genome sequences of CSFV strains. After the RT - PCR method was established and optimized, 15 clinic specimens were detected for CSFV by RT - PCR in comparison with the direct immunofluoresence assay （DFA）. The result indicated that a 509 bp specifical fragment was obtained from the CSFV vaccine strains by RT - PCR with the sensitivity reaching 10 pg of CSFV - RNA. Negative results were obtained from porcine reproductive and respiratory syndrome virus （PRRSV）, porcine parvovirus （PPV）, pseudorabies virus （PRV） and porcine circovirus 11 （PCV2）. The nucleotide sequence homology of RT - PCR products was 95%- 99% of the other CSFV strains. The positive detection rate of the 15 clinic specimens reached 66.7% by RT - PCR, as compared to 60. 0% by DFA. The two method＇s coincidence rate was 80%. These results indicated that the RT- PCR method was more sensitive than DFA, and that both methods were suitable for rapid detection of CSFV.
出处
《福建农业学报》
CAS
2007年第3期231-234,共4页
Fujian Journal of Agricultural Sciences
基金
福建省科技重大专项(2006NZ0003-2)
