关键词: 猪传染性胃肠炎病毒; 猪流行性腹泻病毒; 二重RT-PCR; 检测; 建立; 应用

Abstract: A duplex RT-PCR for detection porcine transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhea virus (PEDV) was developed using the primers designed basing on the N and M gene coding for the nucleoprotein of TGEV and membrane glycoprotein of PEDV respectively. The total RNA of standard TGEV and PEDV strains were used as the positive control to establish the duplex RT-PCR assay. The sensitivity, specificity and repetition assay of duplex RT-PCR were tested, and samples taken from clinic suspicious TGEV or PEDV infected pigs and having been testified by routine cell culture and identification were detected by the established duplex RT-PCR assay. The results indicated that the duplex RT-PCR assay was successfully established. The specificity and sensitivity of the duplex RT-PCR assay revealed that the threshold of duplex RT-PCR was 10 TCID₅₀/mL of TGEV or PEDV, and no products were amplified from the nucleic acid of ST cell or the other 7 kinds of pathogenic viral or bacterial microorganism acting as the negative control. The repetition test indicated that the duplex PCR was repeatable. The detection results for 22 clinic suspicious TGEV or PEDV infected pigs were consistent with the results tested by sequencing. The study suggested that the established duplex RT-PCR method was highly specific and sensitive,and was suitable to clinic rapid identified diagnosing of TGEV and PEDV.

Key words: porcine transmissible gastroenteritis virus; porcine epidemic diarrhoea virus; duplex RT-PCR; detection; establishment; application