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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1994 Oct;32(10):2434–2440. doi: 10.1128/jcm.32.10.2434-2440.1994

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

J T Aubin 1, L Poirel 1, C Robert 1, J M Huraux 1, H Agut 1
PMCID: PMC264080  PMID: 7814479

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.

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Selected References

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  1. Ablashi D. V., Balachandran N., Josephs S. F., Hung C. L., Krueger G. R., Kramarsky B., Salahuddin S. Z., Gallo R. C. Genomic polymorphism, growth properties, and immunologic variations in human herpesvirus-6 isolates. Virology. 1991 Oct;184(2):545–552. doi: 10.1016/0042-6822(91)90424-a. [DOI] [PubMed] [Google Scholar]
  2. Agut H. Puzzles concerning the pathogenicity of human herpesvirus 6. N Engl J Med. 1993 Jul 15;329(3):203–204. doi: 10.1056/NEJM199307153290311. [DOI] [PubMed] [Google Scholar]
  3. Aubin J. T., Agut H., Collandre H., Yamanishi K., Chandran B., Montagnier L., Huraux J. M. Antigenic and genetic differentiation of the two putative types of human herpes virus 6. J Virol Methods. 1993 Feb;41(2):223–234. doi: 10.1016/0166-0934(93)90129-f. [DOI] [PubMed] [Google Scholar]
  4. Aubin J. T., Collandre H., Candotti D., Ingrand D., Rouzioux C., Burgard M., Richard S., Huraux J. M., Agut H. Several groups among human herpesvirus 6 strains can be distinguished by Southern blotting and polymerase chain reaction. J Clin Microbiol. 1991 Feb;29(2):367–372. doi: 10.1128/jcm.29.2.367-372.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bovenzi P., Mirandola P., Secchiero P., Strumia R., Cassai E., Di Luca D. Human herpesvirus 6 (variant A) in Kaposi's sarcoma. Lancet. 1993 May 15;341(8855):1288–1289. doi: 10.1016/0140-6736(93)91198-u. [DOI] [PubMed] [Google Scholar]
  6. Chou S., Marousek G. I. Analysis of interstrain variation in a putative immediate-early region of human herpesvirus 6 DNA and definition of variant-specific sequences. Virology. 1994 Jan;198(1):370–376. doi: 10.1006/viro.1994.1044. [DOI] [PubMed] [Google Scholar]
  7. Collandre H., Aubin J. T., Agut H., Béchet J. M., Montagnier L. Detection of HHV-6 by the polymerase chain reaction. J Virol Methods. 1991 Feb-Mar;31(2-3):171–179. doi: 10.1016/0166-0934(91)90155-s. [DOI] [PubMed] [Google Scholar]
  8. Cone R. W., Huang M. L., Ashley R., Corey L. Human herpesvirus 6 DNA in peripheral blood cells and saliva from immunocompetent individuals. J Clin Microbiol. 1993 May;31(5):1262–1267. doi: 10.1128/jcm.31.5.1262-1267.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dewhurst S., Chandran B., McIntyre K., Schnabel K., Hall C. B. Phenotypic and genetic polymorphisms among human herpesvirus-6 isolates from North American infants. Virology. 1992 Sep;190(1):490–493. doi: 10.1016/0042-6822(92)91240-u. [DOI] [PubMed] [Google Scholar]
  10. Drobyski W. R., Eberle M., Majewski D., Baxter-Lowe L. A. Prevalence of human herpesvirus 6 variant A and B infections in bone marrow transplant recipients as determined by polymerase chain reaction and sequence-specific oligonucleotide probe hybridization. J Clin Microbiol. 1993 Jun;31(6):1515–1520. doi: 10.1128/jcm.31.6.1515-1520.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kondo K., Hayakawa Y., Mori H., Sato S., Kondo T., Takahashi K., Minamishima Y., Takahashi M., Yamanishi K. Detection by polymerase chain reaction amplification of human herpesvirus 6 DNA in peripheral blood of patients with exanthem subitum. J Clin Microbiol. 1990 May;28(5):970–974. doi: 10.1128/jcm.28.5.970-974.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lawrence G. L., Chee M., Craxton M. A., Gompels U. A., Honess R. W., Barrell B. G. Human herpesvirus 6 is closely related to human cytomegalovirus. J Virol. 1990 Jan;64(1):287–299. doi: 10.1128/jvi.64.1.287-299.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Pruksananonda P., Hall C. B., Insel R. A., McIntyre K., Pellett P. E., Long C. E., Schnabel K. C., Pincus P. H., Stamey F. R., Dambaugh T. R. Primary human herpesvirus 6 infection in young children. N Engl J Med. 1992 May 28;326(22):1445–1450. doi: 10.1056/NEJM199205283262201. [DOI] [PubMed] [Google Scholar]
  14. Rajcáni J., Yanagihara R., Godec M. S., Nagle J. W., Kudelova M., Asher D. M. Low-incidence latent infection with variant B or roseola type human herpesvirus 6 in leukocytes of healthy adults. Arch Virol. 1994;134(3-4):357–368. doi: 10.1007/BF01310573. [DOI] [PubMed] [Google Scholar]
  15. Salahuddin S. Z., Ablashi D. V., Markham P. D., Josephs S. F., Sturzenegger S., Kaplan M., Halligan G., Biberfeld P., Wong-Staal F., Kramarsky B. Isolation of a new virus, HBLV, in patients with lymphoproliferative disorders. Science. 1986 Oct 31;234(4776):596–601. doi: 10.1126/science.2876520. [DOI] [PubMed] [Google Scholar]
  16. Schirmer E. C., Wyatt L. S., Yamanishi K., Rodriguez W. J., Frenkel N. Differentiation between two distinct classes of viruses now classified as human herpesvirus 6. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5922–5926. doi: 10.1073/pnas.88.13.5922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wyatt L. S., Balachandran N., Frenkel N. Variations in the replication and antigenic properties of human herpesvirus 6 strains. J Infect Dis. 1990 Oct;162(4):852–857. doi: 10.1093/infdis/162.4.852. [DOI] [PubMed] [Google Scholar]
  18. Yamanishi K., Okuno T., Shiraki K., Takahashi M., Kondo T., Asano Y., Kurata T. Identification of human herpesvirus-6 as a causal agent for exanthem subitum. Lancet. 1988 May 14;1(8594):1065–1067. doi: 10.1016/s0140-6736(88)91893-4. [DOI] [PubMed] [Google Scholar]

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