Real-time fluorescent quantitative PCR assay for detection of human Bocavirus / 临床儿科杂志

ABSTRACT

Objective To establish the method for detecting human Bocavirus (HBoV) by a real-time fluorescent quantitative PCR (FQ-PCR) and to assay HBoV from nasopharyngeal samples of children with acute respiratory tract infections.Methods The specific primers and Taqman probes of targeted HBoV NP1 gene were designed.The aimed fragment of NP1 gene was amplified with PCR and ligated into a pCEM-T Easy vector to set up the standards.The method for detection of NP1 gene using FQ-PCR was established.The sensitivity and specificity of the method had been tested.Then 195 clinical specimens were subsequently tested.Results The specificity of this method was excellent.The linear amplification of the assay ranges from 10 copies/μl to 10~8 copies/μl.Twelve specimens were tested positive for HBoV in 195 clinical specimens (6.2%).Conclusions The method for detection of HBoV NP1 geneby real-time fluorescent quantitative PCR was established successfully.It can offer fast and reliable diagnosis for HBoV infection.