犬细小病毒的PCR诊断试剂盒的研制

Development of the Polymerase Chain Reaction Diagnostic Kit of Canine Parvovirus DNA

目的 建立检测犬细小病毒的PCR诊断试剂盒。方法 通过在犬细小病毒 (CPV)的基因组中设计的特异性寡核苷酸引物 ,利用PCR技术研制犬细小病毒的PCR诊断试剂盒。结果 在特定的反应条件下 ,使用该试剂盒能特异地扩增含有犬细小病毒的样品 ,且能检出痕量 (10ng)的犬细小病毒DNA ,被测粪样只需进行简单煮沸就能进行PCR反应 ,该试剂盒能在常温下保存 1周以上、4℃保存 1年以上。同血凝试验 (HA〕及单克隆抗体ELISA方法比较 ,对 6 6份样品进行检测 ,显示该PCR试剂盒具有特异、灵敏等优点。结论 成功研制了犬细小病毒的PCR诊断试剂盒 。

Objective To develop a polymerase chain reaction(PCR) diagnostic kit for the detection of canine parvovirus(CPV). Methods\ A polymerase chain reaction technology was used. Results\ To increase the sensitivity and specificity of the reaction, a double specific primer was designed in the CPV VP1/VP2 gene. 10 ng of viral replicative form(RF) DNA could be detected by the CPV PCR Kit. CPV DNA from feces could be enabled for the test after a fast and simple boiling pretreatment. The PCR assay was more sensitive than HA assay and McAbs ELISA. CPV PCR Kit stored in 4℃ would stay active for one year. Conclusion\ CPV PCR Kit was developed successfully, and it could be an alternative technique for routine parovirus diagnosis.