摘要
为了实现猪细环病毒(TTSuV)快速检测,给TTSuV的调查、监测和防控提供可行性、操作性强的技术支持。本试验根据TTSuV的序列特点设计特异性引物、探针,应用实时荧光PCR技术检测TTSuV1a型和TTSuV1a1b型。以猪细小病毒、猪圆环病毒Ⅱ型和猪伪狂犬病毒进行特异性试验,无交叉反应。该检测方法具有较好的特异性和重复性,TTSuV1a灵敏度可达9.2拷贝/μL,TTSuV1b灵敏度可达5.6×101拷贝/μL;批内变异系数均小于1%,批间变异系数均小于3%。将该方法与PCR法、PCR-DHPLC进行比较,结果证实该方法具有较好的准确性。对92份血清样本进行检测,发现有58份TTSuV1a阳性、62份TTSuV1b阳性、51份TTSuV1a和TTSuV1b共感染阳性。本研究建立的TTSuV实时荧光PCR法具有特异、敏感、快速、重复性好等优点,可用于TTSuV感染的分子流行病学调查。
To identify the Torque teno sus virus（TTSuV）,real-time PCR assay was performed in this study. Specific primers for the partial region of the TTSuVla and TTSuVlb were selected to conduct the real-time PCR assays. The specificity test was performed with PPV, there was not cross reaction between PCV Ⅱ and PRV, and better specificity and repeatability were abserved. Sensitivity analysis showed that the developed real-time PCR for TTSuVla could detect 9.2 cop- ies/μL,and 5. 6 × 101 copies/μL for TTSuVlb. The coefficient of variation in a group was less than l~,the coefficient of variation between the groups was less than 3%,with better accuracy compared with the PCR,PCR-DHPLC. To detect the serum samples, 58 TTSuVla positives re- suits could be observed by real-time PCR for 92 samples,62 TTSuVlb positives,51 TTSuVla and TTSuVlb coinfected positives. The method with good specification, sensitivity, repeatability, and quickness could be used for epidemiological investigation of TTSuV.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2017年第6期999-1006,共8页
Chinese Journal of Veterinary Science
基金
江西省科技计划资助项目(20151BBF60048)
