title: Determination of the six major human herpesviruses in cerebrospinal fluid and blood specimens of children.
abstract: OBJECTIVE
  To detect and differentiate six major human herpesviruses in the cerebrospinal fluid (CSF) and blood of children by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP).
  
  
  METHODS
  We synthesized two pairs of primers in the well-conserved regions of the DNA polymerase gene in human herpesviruses. One pair was designed to amplify cytomegalovirus (CMV), Epstein-Barr virus (EBV), herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), and the other pair to amplify varicella-zoster virus (VZV) and human herpesvirus 6 (HHV-6) by PCR. Virus species identification was achieved by restriction enzyme digestion with BamHI and BstUI. Ninety-eight CSF and 75 blood specimens were analysed by this technique. At the same time, all blood specimens were also examined by enzyme-linked immunosorbent assay (ELISA).
  
  
  RESULTS
  Thirteen (13.3%) of 98 CSF specimens and 26 (34.7%) of 75 blood specimens were positive for herpesvirus DNA in this PCR assay. Only 10 (13.3%) of the blood specimens were positive in ELISA for virus-IgM antibody. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PCR in detecting herpesvirus infections compared with ELISA were 100% (10/10), 75.4% (49/65), 38.5% (10/26) and 100% (49/49), respectively. These results indicate that the positive rate of PCR was significantly higher than that of ELISA (p < 0.05). The herpesvirus type of these positive specimens was rapidly detected using restriction enzyme digestion with BamHI and BstUI.
  
  
  CONCLUSIONS
  PCR-RFLP is a specific, sensitive and accurate technique for the identification of herpesvirus infections in the CSF and blood of children.
authors.affiliation: Department of Infectious Medicine, Children's Hospital of Zhejiang University, School of Medicine, Hangzhou, China. dongguanp@yahoo.com.cn
authors.forename: G
authors.initials: G
authors.lastname: Dong
authors.forename: S
authors.initials: S
authors.lastname: Shang
authors.forename: L
authors.initials: L
authors.lastname: Liang
authors.forename: X
authors.initials: X
authors.lastname: Yu
mesh_terms: D018141:Betaherpesvirinae / Q000235:genetics / Q000302:isolation & purification*; D020805:Central Nervous System Viral Diseases / Q000097:blood* / Q000134:cerebrospinal fluid*; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D014645:Herpesvirus 3, Human / Q000235:genetics / Q000302:isolation & purification*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase