Amplification and detection of the six major Heperviruses by PCR-based assay
- VernacularTitle:体外6种疱疹类病毒DNA扩增及其基因分型
- Author:
Guanping DONG
;
Juanfen LU
;
Shiqiang SHANG
- Publication Type:Journal Article
- Keywords:
DNA-directed DNA polymerse;
Polymorphism, restrictionfragment length;
Cloning, molecular
- From:
Chinese Journal of Infectious Diseases
1999;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To detect and differentiate six major human herpesviruses DNA by PCR, RFLP, DNA clone and sequence analysis. Methods Based on the sequence of well coonserved regions of the DNA polymerse gene in human herpesvimses, we synthesized two pairs of primers, including one pair designed to amplify herpes simplex virus type 1 and 2, Epstein Barr virus and cytomegalovirus, other pair of primer to varicella zoster virus and human herpesvirus 6 by PCR. Identification of the virus species was achieved through restriction enzyme digestion with BamHI and BstUI. Results The products of six human herpesviruses after PCR amplification were from 510bp to 592bp and allowed characterization of herpesvirus type with restriction endonulease analysis. The sensitivity could reach 0.1fg DNA and had no cross reaction to human genomic DNA, bacteria, fungas and virus. 89 cerebrospinal fluids(CSF) and 75 blood specimens were tested for the presence of hepersvirus DNA by this PCR assay, in which 13(14.6%) CSF and 26(34.7%) blood specimens were positive. Comparatively, 10(13.3%) were positive by ELISA for HSVⅠ/Ⅱ,EBV,CMV IgM in blood specimens and significantly lower than that of the PCR rate (P
