Polymerase chain reaction amplification and in situ hybridization for the detection of human B-lymphotropic virus.
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.
Buchbinder
A
A
Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, Maryland 20892.
Josephs
S F
SF
Ablashi
D
D
Salahuddin
S Z
SZ
Klotman
M E
ME
Manak
M
M
Krueger
G R
GR
Wong-Staal
F
F
Gallo
R C
RC
D000163:Acquired Immunodeficiency Syndrome / Q000382:microbiology; D002051:Burkitt Lymphoma / Q000382:microbiology; D002460:Cell Line; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D005784:Gene Amplification; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008232:Lymphoproliferative Disorders / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D015347:RNA Probes; D012367:RNA, Viral / Q000032:analysis
D015342:DNA Probes; D004279:DNA, Viral; D015347:RNA Probes; D012367:RNA, Viral