鸭圆环病毒巢式PCR检测方法的建立及应用

鸭圆环病毒病给养鸭业造成严重的经济损失,鉴于此,本研究根据GenBank登录的鸭圆环病毒(duck circovirus,DuCV)保守区基因组序列(登录号:NC_005053.1),设计合成内、外2对引物,通过PCR反应条件的优化,建立了检测DuCV的巢式PCR检测方法。该方法对鸭病毒性肝炎病毒、鸭瘟病毒、鸭细小病毒、新城疫病毒的扩增结果均为阴性;第1轮扩增的灵敏度为10 pg,第2轮扩增的灵敏度为0.1 pg,通过巢式PCR,敏感性提高了100倍。本研究建立的巢式PCR方法具有特异性强、敏感性高、重复性好等优点,可以准确快速的用于鸭圆环病毒的病原检测。

Establishment and Application of Nest PCR Method for Detection of Duck Circovirus

Because duck circovirus (DuCV) caused severe economic losses,based on DuCV conserved region in GenBank, two pairs of primers were designed and synthesis. Through optimization of PCR reaction conditions, a nested PCR method had been developed for detection of duck circovirus. Amplification results of duck viral hepatitis virus, duck plague virus, duck parvovirus, Newcastle disease virus were negative using the established method. The first amplification sensitivity was 10 pg, and the second amplified sensitivity was 0.1 pg, sensitivity was increased by 100 times by nested PCR. The established nested PCR method had strong specificity, high sensitivity, good repeatability and could be used rapidly and accurately for duck circovirus pathogen detection.