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Background: This study was undertaken to establish a restriction endonulease pattern which could detect and differentiate four major human herpesviruses by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP), and to compare PCR-RFLP with enzyme-linked immunosorbent assay (ELISA) in diagnosing herpesvirus infection.
Methods: A pair of primers was designed to amplify herpes simplex virus type 1 and 2 (HSV-1/-2), Epstein-Barr virus (EBV) and cytomegalovirus (CMV). At last, we used the PCR-RFLP technique to differentiate four different herpesviruses. Meanwhile, 75 clinical blood specimens from infants of suspected viral infection and 38 blood specimens from healthy children were evaluated for herpesviruses DNA by PCR-RFLP or virus-specific IgM antibody detection by ELISA.
Results: The products of four human herpesviruses after PCR amplification varied from 510 bp to 592 bp and allowed characterization of herpesvirus type with restriction endonulease analysis. Among the 75 specimens, 23 (30.7%) were shown positive by PCR including 13 for CMV, 4 for EBV, 5 for HSV-2, and 1 for HSV-1 after restriction enzyme digestion with BamHI and BstUI, whereas 10 (13.3%) were detected positive by ELISA. All ELISA-positive specimens were likewise positive by PCR. Thirteen of 65 ELISA-negative specimens were tested positive by PCR.
Conclusion: The PCR-RFLP technique is more specific, sensitive, rapid and accurate than ELISA in diagnosing herpesvirus infection.
Key words: herpesvirus; polymerase chain reaction; restriction fragment length; polymorphism; ELISA
World J Pediatr 2007;3(2):129-133
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