Development of SYBR Green real-time PCR assay for detection of adenovirus

Objective To develop a rapid SYBR Green real-tmie PCR a ssay for early detection of human adenovirus and quantification of the virus nucleic acid.Methods The amplification system and the rmal cycle condition of the SYBR Green real-tmie PCR were developed and optmiized.Specificity,sensitivity and reproducibility of the method were evaluated.The characteristic T_m of the target was determinded by preforming a melting curve analysis on the PCR product.The standard curve and virus quantitative analysis model were constructed.Eighty-fourc linical specmiens were detected with the method.Results No cross-amplification was observed when other respiratory virus was tested.As few as 10² copies/μl plasmids were reliably detected by the method.Melting curve analysis showed a single melting peak with a T_m of 84.9±0.1℃.A deno virus DNA was detected in 30.95% of the clinical specmiens.Conclusion Arapid,sensitive and specific SYBR Green real-tmie PCR for analysis of adenovirus DNA was established which was suitable for early diagnosis and virus quantification.