【摘要】 目的在省级脊髓灰质炎(脊灰)实验室建立一种快速、灵敏、准确的脊灰病毒(Poliovirus,PV)型内鉴定(Intratypic Differentiation,ITD)及基因型鉴定的方法。方法以PV衣壳蛋白(Capsid Protein)VP1编码区基因序列为目标,设计并合成引物和Taqman探针,建立实时荧光定量RT-PCR(Real-time RT-PCR,r RT-PCR)检测体系,并考察该方法的重复性、灵敏性和特异性。结果该方法能快速、灵敏地鉴定出PV血清型及毒株类型,在1.0×108copies/μl~1.0×103copies/μl检测范围之间有良好的线性关系,相关系数为0.993,最低检测限为103.5CCID50/0.1 ml。结论成功在省级实验室建立了PV的r RT-PCR检测技术,该技术特异性强,敏感性高,操作简便快速,适用于PV的型内鉴定和基因型鉴定,可应用于实验室诊断,为免疫策略快速提供依据。更多还原

【Abstract】 Objective To establish a rapid, sensitive and accurate identification for poliovirus(PV) in provincial which can Intratypic Differentiation(ITD) and identify genotype. Methods We designed and synthesized of primers and Taqman probe by using VP1 coding region of PV capsid protein as the target and establish a Real-time RT-PCR detection system. At the same time, we studied the sensitivity, specificity and reproducibility of the method. Results Real-time RT-PCR can identify the serotype and genotype of PV sensitively and quickly, and there was a good linear relationship between 1.0×108copies/μl~1.0×103copies/μl detection range. The correlation coefficient is 0.993, and the minimum detection is 103.5CCID50/0.1ml. Conclusion We established Real-time RT-PCR PV detect technology successfully in provincial laboratory, and that technology was with strong specificity, high sensitivity and simple, fast operation, so it was suitable for Intratypic Differentiation(ITD) and identify genotype of PV and it could be used in laboratory diagnosis and provided the basis for immunization strategies.更多还原