Determination of Adenovirus Type 7 by Cells Culture Combined with RT-PCR
- VernacularTitle:细胞培养结合实时荧光RT-PCR法快速检测7型腺病毒感染
- Author:
Dingkun SHANG
;
Baoling SHEN
;
Xiaoqun ZHENG
;
Ying PENG
;
Xinxin LIN
- Publication Type:Journal Article
- Keywords:
Adenovirus;
Cell culture;
Real-time;
RT-PCR
- From:
Chinese Journal of Nosocomiology
1994;0(04):-
- CountryChina
- Language:Chinese
-
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.
