Determination of six major human herpes viruses in cerebrospinal fluid and blood of children with consensus primers.
- Author:
Guan-ping DONG
1
;
Shi-qiang SHANG
;
Zhong-sheng YU
;
Li LIANG
;
Xi-lin YU
Author Information
- Publication Type:Journal Article
- MeSH: Child; Cytomegalovirus; isolation & purification; Cytomegalovirus Infections; virology; DNA Primers; DNA, Viral; blood; cerebrospinal fluid; Epstein-Barr Virus Infections; virology; Female; Herpesviridae; isolation & purification; Herpesviridae Infections; virology; Herpesvirus 4, Human; isolation & purification; Humans; Male; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Simplexvirus; isolation & purification
- From: Journal of Zhejiang University. Medical sciences 2005;34(1):60-64
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo identify 6 major human herpesviruses with consensus primers and to explore its clinical application.
METHODSBased on the highly-homogeneous regions of DNA polymerase gene in human herpesviruses,Two pairs of primer were synthesized. One pair was designed to amplify herpes simplex virus type 1, type 2, Epstein-Barr virus and cytomegalovirus; and another was used to amplify varicella-zoster virus or human herpesvirus 6. Virus species identification was performed by restriction enzyme digestion with BamH I and BstU I. Thirty-eight CSF specimens of clinically diagnosed viral encephalitis,and 49 blood specimens from 27 confirmed cases and 22 clinically diagnosed ones were tested for herpes virus DNA using the PCR-RFLP assay with these primers.
RESULTSThirteen out of 38 CSF specimens (34.2%) were herpes virus positive. All blood specimens from 27 confirmed cases showed positive results, while for 22 clinically diagnosed cases 16 (72.7%) were positive. The types of herpes virus were determined using restriction enzyme digestion with BamH I and BstU I. Two CSF specimens from the patients, who were treated with aciclovir for 2 - 3 days, were still positive for herpes virus DNA by this method. None of the control blood or CSF controls were positive for herpesvirus by PCR.
CONCLUSIONThe PCR-RFLP method used in this study is a specific, sensitive and practicable one for diagnosis of herpes virus infection.