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 wellcharacterized 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.

Human herpesvirus‐6 infection: A prospective study evaluating HHV‐6 DNA levels in liver from children with acute liver failure

“…Samples were considered positive when at least one copy was detected in the reactive well. The identification of the variant of HHV-6 (A or B) was carried out using a primer-specific qualitative PCR, as previously described [Aubin et al, 1994]. The detection of HCMV DNA was also performed on the same biological samples by means of qualitative PCR, as previously described [Drouet et al, 1993].…”

Variability of gB and gH genes of human herpesvirus‐6 among clinical specimens

“…The origin of four HHV-6 variant A strains (SIE, TAN, GS, and U1102) and seven HHV-6 variant B (HST, BLA, BLE, BOB, BOU, MAR, and MBE) strains studied have been described previously [Agut et al, 1988;Aubin et al, 1991Aubin et al, , 1994. These strains were propagated in phytohaemagglutinin-stimulated peripheral blood mononuclear cells (PBMCs) issued from healthy donors, except for GS and HST which were propagated in HSB-2 and MT4 cell lines, respectively.…”

Prevalence of viral infection markers by polymerase chain reaction amplification and interferon‐alpha measurements among patients undergoing lumbar puncture in an emergency department

“…A 267 bp long fragment from the XbaI M region of VZV genome was amplified and then hybridized using a PCR assay published previously [Puchhammer-Stöckl et al, 1991]. HHV-6 amplification of a 350 bp long fragment in the major capsid protein gene was carried out and then hybridized [Aubin et al, 1994]. CMV DNA was detected until May 2001 using a conventional PCR assay [Gozlan et al, 1992] and since June 2001 with a TaqMan 1 (PE Biosystems, Cheshire, UK) PCR assay [Najioullah et al, 2001].…”