Polymerase chain reaction amplification and in situ hybridization for the detection of human B-lymphotropic virus

J Virol Methods. 1988 Sep;21(1-4):191-7. doi: 10.1016/0166-0934(88)90065-1.

Abstract

Polymerase chain reaction amplification (PCR) is a recently described technique that allows for the amplification of a given sequence of DNA. It can be used to reliably amplify sequences of up to 3 kb within hours. The amplified sequence can then be recognized by hybridization with a specific probe after transfer onto nitrocellulose or nylon paper. We used PCR to recognize human B-lymphotropic virus (HBLV or HHV-6) specific sequences in various tumors as well as in the blood of patients with AIDS. Sixty-three specimens of DNA extracted from peripheral blood of patients with AIDS as well as DNA extracted from various lymphoproliferative disorders were analysed; 52 out of 63 (83%) patients with AIDS were found to have amplification of the HHV-6 specific sequence; 2 out of the 63 (3%) had equivocal amplification and 9 (14%) were found to be negative. Twenty out of 23 tumors were found to have amplified HBLV-specific sequences. Only one of these tumors was positive by Southern hybridization on restriction enzyme digested genomic DNA. In situ hybridization of clinical specimens using radiolabelled RNA probes or hapten-labelled DNA probes was used to detect the presence of HBLV in tumors. Three tumors of B cell origin were found to be positive for HBLV.

MeSH terms

  • Acquired Immunodeficiency Syndrome / microbiology
  • Burkitt Lymphoma / microbiology
  • Cell Line
  • DNA Probes
  • DNA, Viral / analysis*
  • Gene Amplification*
  • Herpesviridae / genetics
  • Herpesviridae / isolation & purification*
  • Humans
  • Lymphoproliferative Disorders / microbiology
  • Nucleic Acid Hybridization*
  • RNA Probes
  • RNA, Viral / analysis

Substances

  • DNA Probes
  • DNA, Viral
  • RNA Probes
  • RNA, Viral