Detection of Borna disease virus by fluorescence quantitative nested RT-PCR
- VernacularTitle:博尔纳病病毒荧光定量套式RT-PCR检测方法的建立
- Author:
Ping XU
- Publication Type:Journal Article
- Keywords:
Borna disease virus;
Genes;
Viral;
Polymerase chain reaction
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To establish fluorescence quantitative nested RT-PCR method for detecting Borna disease virus (BDV).Methods:According to the specific sequence of BDV P24 genes,the primers and the fluorescence probe were designed and synthesized.The fragment generated by PCR was cloned into the pMD18-T vector.The positive recombinant plasmid would be used as standard quantitative template to make the standard curve for sample detection.Results:The standard curve indicated the linear relationship between Ct(cycle threshold) and template concentration (r=0.998).The fluorescence quantitative nested RT-PCR method for detection of BDV p24 fragment was established.And it was used to detect BDV p24 fragment in peripheral blood mononuclear cells(PBMC)from 58 patients with neuropsychiatric disorders and 50 healthy blood donors.4 patients with neuropsychiatric disorders were positive,and normal control negative.Conclusion:The fluorescence quantitative nested RT-PCR method for detection of Borna disease virus can eliminate PCR cross-contamination which causes false positive,and real-time detection ensures accurate quantity.It can be used to study the association between BDV infection and human neuropsychiatric disorders.
