Differential diagnosis of HTLV-I and HTLV-II infections by restriction enzyme analysis of 'nested' PCR products.
A 'nested' polymerase chain reaction (PCR) assay is described which is capable of detecting single copies of human T-cell lymphotropic virus (HTLV) in genomic DNA extracted from peripheral blood mononuclear cells (PBMCs). A single set of 'nested' oligonucleotide primers, based on the highly conserved tax/rex region of the viral genome, was able to detect both HTLV-I and HTLV-II proviral sequences in clinical samples of diverse geographical origins, from the United States, Great Britain, Japan, the Caribbean, Italy, Greece, Iraq and West Africa. Rapid discrimination between HTLV-I and HTLV-II infections was achieved by restriction enzyme analysis of unpurified second-round PCR products, even in those cases in which serological assays had failed to provide a definitive result. Over a 2-year period, a total of 53 HTLV infections (37 HTLV-I and 16 HTLV-II) were identified by this technique and complete concordance with serological typing, available in 41 cases, was observed.
Tuke
P W
PW
Department of Medical Microbiology, University College and Middlesex School of Medicine, London, UK.
Luton
P
P
Garson
J A
JA
D001483:Base Sequence; D002460:Cell Line; D004279:DNA, Viral / Q000032:analysis; D015252:Deoxyribonucleases, Type II Site-Specific; D003937:Diagnosis, Differential; D015490:HTLV-I Infections / Q000175:diagnosis* / Q000382:microbiology; D015491:HTLV-II Infections / Q000175:diagnosis* / Q000382:microbiology; D015368:Human T-lymphotropic virus 1 / Q000235:genetics / Q000302:isolation & purification; D015367:Human T-lymphotropic virus 2 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000175:diagnosis / Q000382:microbiology; D007963:Leukocytes, Mononuclear / Q000382:microbiology; D008969:Molecular Sequence Data; D015493:Paraparesis, Tropical Spastic / Q000175:diagnosis / Q000382:microbiology; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012703:Serotyping
D004279:DNA, Viral; D015252:Deoxyribonucleases, Type II Site-Specific; C024405:GATC-specific type II deoxyribonucleases; C027046:TCGA-specific type II deoxyribonucleases