Identification of human herpesvirus 6 variants A and B by amplimer hybridization with variant-specific oligonucleotides and amplification with variant-specific primers

Abstract

Two distinct PCR-based procedures were evaluated for the detection and identification of human herpesvirus 6 (HHV-6) variants A and B in uncultured human samples. Variant-specific oligonucleotide hybridization (VSOH) is based on the amplification of two distinct regions of the HHV-6 genome, followed by hybridization of amplimers with variant-specific oligonucleotide probes. Variant-specific primer PCR (VSPP) is based on the amplification of each variant by using variant-specific primers. The study of 10 well-characterized HHV-6 strains allowed us to demonstrate the high sensitivity and specificity of both methods. With variant mixtures, however, some limitations of VSOH were evidenced and VSPP was required to obtain unambiguous results. The combination of VSOH and VSPP was applied to the direct study of 300 peripheral blood mononuclear cell samples from French subjects. HHV-6 was detected in 15 samples: 11 corresponded to variant B, 3 corresponded to variant A, and 1 corresponded to a mixture of both variants.