Application of TaqMan based real time fluorescent quantitative RT-PCR assay in rapid detection of enteroviruses

Objective To establish a rapid,sensitive and specific TaqMan RT-PCR method to detect the enteroviruses.Methods Specific primers and TaqMan probes were designed based on the alignment results of the 5′ untranslated region(5′-UTR) of the enteroviruses.The reaction systems and conditions were optimized.The sensitivity,specificity and repeatability of the method were validated.And the practical application of the method for the clinical samples was evaluated Results TaqMan RT-PCR was specific to detect the polioviruses type Ⅰ,Ⅱand Ⅲ,the cosaxieviruses and eochviruses.But no reaction was observed with the hepatitis A virus,Japanese encephalitis virus,Dengue virus,adenovirus,Herpes simplex virus and norovirus.The detection limit was 10-1TCID50/100 μl.Among the 12 conjunctiva swab samples from the acute conjunctivitis patients,there were 7 swabs positive detected by TaqMan RT-PCR,while there were 5 swabs positive by general RT-PCR.Conclusion TaqMan RT-PCR is a rapid,sensitive and specific method and could be used to detect the enteroviruses in laboratory.