鸡传染性腺胃炎相关的圆圈病毒3型SYBR GreenⅠ实时荧光定量PCR方法的建立

Development of A SYBR Green Ⅰ Real-time Fluorescent Quantitative PCR Method for Detection of Gyrovirus 3 of Chickens

本研究针对病毒基因组5'端非编码区的保守区域设计引物,建立了用于检测鸡传染性腺胃炎相关的圆圈病毒3型(Gyv3)的SYBR Green Ⅰ荧光定量PCR方法。该方法具有较好的特异性,敏感性和重复性,对于鸡Gyv3以外的其他常见鸡病毒性病原的DNA或cDNA均无特异性扩增,灵敏度能达到30.76 copies/μL,组内和组间变异系数均不超过2%。对临床样品的检测结果显示,本研究建立的荧光定量PCR方法对鸡Gyv3检出率为12.28%。

Gyrovirus 3(GyV3)has been demonstrated to be involved in transmissible viral proventriculitis of chickens.In this study,a SYBR Green Ⅰ real-time PCR was developed for detection of GyV3 infection in chickens by using a pair of primers designed from the 5'untranslated region of genome.The method had the advantage of specificity,sensitivity and repeatability.The PCR method had no amplifications for the DNA or cDNA of other chicken viral pathogens.The detection limit of GyV-plasmid was 30.76 copies/μL and the variation coefficients of intra-assays and inter-assays were both less than 2%.The test results of the clinical samples showed 12.28% positive rate of GyV3 infection.