Determination of Adenovirus Type 7 by Cells Culture Combined with RT-PCR / 中华医院感染学杂志

ABSTRACT

OBJECTIVE To establish a method for detecting adenovirus type 7 by cells culture combined with real-time fluorescent RT-PCR.METHODS After purified adenovirus was dissociated from nasopharyngeal secretion in A549 cells,ADV7 E1A genes were detected by real-time RT-PCR assay and sequence analysis of cells infected with 0.1,0.5,5.0 and 10.0 MOI ADV7 at 3,6,12 and 24 h postinfection.Then the adenovirus in nasopharyngeal secretion was detected with the similar method.RESULTS Early transcription of E1A genes of adenovirus type 7 could be detected by real-time RT-PCR at 3 h postinfection with 0.5MOI virus;or at 6 h postinfection with 0.1MOI virus;Early transcription of E1A genes could be detected at 6 h postinfection in nasopharyngeal secretion.CONCLUSIONS The method by cells culture combined with real-time fluorescent RT-PCR is sensitive,specific and rapid.It can be applied in clinics for diagnosis of adenovirus type 7 infection.