应用逆转录套式PCR检测猪流行性腹泻病毒

摘要

Based on the published nucleotide sequence of porcine epidemic diarrhea virus N gene, two pairs of primers were designed and synthesized, which outer primers amplified fragment length was 297 bp.and inner primers amplified fragment length was 212 bp. RT-nested PCR methods had been established through optimization of the RT-PCR reaction conditions, this method was more sensitive and reliable than ordinary RT-PCR, and could effectively reduce the false positive and pollution. It could be used for rapid diagnosis and epidemiological investigation of the PEDV.%根据已发表的猪流行性腹泻病毒(PEDV)N基因核苷酸序列,设计合成2对引物,其中外引物扩增片段长度为297 bp,内引物扩增片段长度为212 bp,在优化RT-PCR反应条件的基础上,建立了检测PEDV的套式RT-PCR检测方法,该方法较普通RT-PCR更加灵敏、可靠,能有效降低假阳性和污染,可用于PEDV的快速诊断和流行病学调查.