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根据基因库中鸭瘟病毒强毒株和弱毒株UL2基因保守区设计特异性引物,优化PCR反应的引物浓度和退火温度等,初步建立了可同时鉴别鸭瘟病毒强毒株和弱毒株的PCR检测方法,并对建立的方法进行了敏感性、特异性验证和临床样品检测。该PCR检测方法最低能检出1pg的鸭瘟病毒强毒和弱毒DNA模板。对鸭瘟病毒强毒和弱毒模板的检测,得到了与试验设计相符的827bp(强毒)和299bp(弱毒)的扩增条带,而对鸭副黏病毒、鸭坦布苏病毒、鸭圆环病毒、番鸭细小病毒、鸭Ⅰ型肝炎病毒、禽流感病毒和小鹅瘟病毒等病原体的检测均为阴性。说明建立了一种敏感性高、特异性好的鉴别鸭瘟病毒强毒和弱毒的PCR检测方法。
According to the primers designed according to the conserved region of virulent and attenuated strains of duck plague in the gene bank and the primer concentration and annealing temperature of the PCR reaction, the PCR for the simultaneous identification of the virulent and attenuated strains of duck plague Detection methods, and the established method of sensitivity, specificity and clinical sample testing. The PCR detection method can detect 1 pg of duck virulent virulent and attenuated DNA template. The virulent and virulent templates of duck plague virus were tested. The amplified bands of 827bp (virulent) and 299bp (attenuated virulent) were obtained in accordance with the experimental design, while the amplification of duck paramyxovirus, The detection of pathogens such as circovirus, Muscovy duck parvovirus, duck hepatitis A virus, avian influenza virus and gosling pestivirus were all negative. This indicated that a sensitive and specific PCR assay for identifying virulent and attenuated plague virus was established.