二温式多重PCR检测对虾的白斑综合征病毒和桃拉综合征病毒

根据多重PCR的技术原理,利用对虾(Penaeus vannamei)白斑综合征病毒和桃拉综合征病毒的基因序列分别设计了两对特异引物,并将常规三温式PCR扩增程序简化为2个温度梯度,建立二温式多重PCR技术用于对虾白斑综合征病毒(White spot syndrome virus,WSSV)和桃拉综合征病毒(Taura syndrome virus,TSV)的复合快速检测。利用二温式PCR能特异地扩增出WSSV和TSV的基因片段。结果表明,二温式多重PCR技术具有较高的特异性和敏感性,最低能检测到WSSV核酸模板10pg,TSV核酸模板100pg,且对其它一些对虾病原呈阴性。

Two-temperature and Multiple PCR for the Simultaneous Detection of White spot syndrome virus and Taura syndrome virus in Penaeus vannamei

Based on the principle of the multiple PCR, two sets of specific primers were designed according to the gene sequences of White spot syndrome virus ( WSSV ) and Taura syndrome virus ( TSV ) to detect WSSV and TSV simultaneously. In the process of detection, a new modified two-temperature and multiple PCR was developed from three temperature and regular PCR. It showed that all samples which contained WSSV and TSV could be simultaneously detected by the two-temperature and multiple PCR, and no specific band was amplified from control group of vibrio pathogen separated from shrimp Penaeus vannamei. By this modified PCR methool, the sensitivity for detection of WSSV and TSV were 10 and 100 pg, respectively. It indicated that this two-temperature and multiple PCR had high specificity and sensitivity.