Determination of six major human herpes viruses in cerebrospinal fluid and blood of children with consensus primers / 浙江大学学报·医学版

ABSTRACT

<p><b>OBJECTIVE</b>To identify 6 major human herpesviruses with consensus primers and to explore its clinical application.</p><p><b>METHODS</b>Based 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.</p><p><b>RESULTS</b>Thirteen 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.</p><p><b>CONCLUSION</b>The PCR-RFLP method used in this study is a specific, sensitive and practicable one for diagnosis of herpes virus infection.</p>