Determination of the six major human herpesviruses in cerebrospinal fluid and blood specimens of children.
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.
Dong
G
G
Department of Infectious Medicine, Children's Hospital of Zhejiang University, School of Medicine, Hangzhou, China. dongguanp@yahoo.com.cn
Shang
S
S
Liang
L
L
Yu
X
X
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*
D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase