[Development of a polymerase chain reaction (PCR) system for detection of Epstein-Barr virus 1,2--application to...

A one-step PCR system was developed for detection of two types of EBV, using a single pair of primers that were complementary to both sequences coding the EBV nuclear antigen (EBNA)-2, and encompassing a large deletion in the sequence of type 1 EBV. Type 1 and type 2 EBV were detected simultaneously by polyacrylamide gel electrophoresis and ethidium bromide stains after amplification. The specificity of amplification was confirmed by Southern blot hybridization with the oligonucleotide probe that was complementary to both EBNA-2 regions. Additionally, two pairs of type specific primers were synthesized from divergent sequences in the EBNA-2 regions of type 1 and type 2 EBV. The PCRs were employed for analysis of the EBV genotype in the oral cavities of healthy donors and patients with various tonsillar disorders. EBV was detected in 60 samples from healthy donors and patients with various tonsillar disorders. Fifty-six contained type 1 and four type 2. Double infection was not seen in either healthy donors or patients. These results indicate that type 1 EBV is highly dominant in Japan. It is interesting to note that patients with acute tonsillitis and palmoplantar pustulosis showed a statistically higher frequency (P < 0.01) of EBV excretion than healthy donors. In examining anti-EBV viral capsid (VCA)-IgG antibody titers and EBV excretion in the oral cavity, EBV excretion was not found to correlate with the VCA-IgG antibody titers.