人3型腺病毒荧光定量PCR检测方法的建立

Detection of Human Adenovirus Type-3 with Real Time PCR

目的通过建立人3型腺病毒荧光定量PCR检测方法，快速鉴定人3型腺病毒感染，为早期诊断提供依据。方法以人3型腺病毒感染的细胞和常见的呼吸道消化道感染的病原体为研究对象，根据病毒六邻体高度保守的序列，设计荧光定量PCR反应体系，并分析实验的敏感性和特异性。结果人3型腺病毒TCID50细胞培养液中病毒核酸最低检出稀释度是10^-6，对应的拷贝数分别是5．802×10^3copies／mL和6．968×10^2copies／mL．呼吸道和消化道感染常见的病原体未出现交叉反应。结论荧光定量PCR的应用，可有效缩短检测时间，提高检测的敏感度。

Objective To establish a rapid real time PCR （RT-PCR） diagnostic method for human adenovirus type-3 virus. Methods Cells infected by human adenovirus type-3 virus and other common respiratory and digestive tract pathogens were used in the study. The highly conserved sequences were used in the design of probes for the RT- PCR. Sensitivity and specificity were the analysed. Results The detection limit of human adenovirus type-3 virus in the infected cell culture medium was 10-6-fold of dilution. The correspording copies were 5.862×10^3 copies/mL and 6.968×10^2 copies/mL respectively.The probes were highly specific for the human adenovirus type-3 virus. Conclusion The developed RT-PCR method can be used to improve the detection sensitivity and shorten the detection time.