Therefore, a rapid, specific, and sensitive method is required for detecting the HTLV-I viral genome. Southern-blot analysis is relatively time-consuming and re- quires a considerable amount of DNA. Genomes, clonality of the infected cells during proliferation, and variation of the integrated viral genomic DNA (12,13). The Southern-blot analysis provides some information about infected cells, such as the number of integrated viral Therefore, the potential transmission of HTLV-I from asymptomatic, sero-negative individuals necessitates the development of a sensitive assay for the viral genome.

For ex- ample, virus particles were barely detected, even in ATLA- positive cultures ( 1 I ). However, im- munological assays directed to viral protein antigens may not detect the virus during certain stages of infection. Furthermore, the possibility of vertical trans- mission via the carrier mothers milk has also been sug- gested (6).Р вЂќР вЂћssays developed for the detection of antibodies to HTLV- I proteins-ATL-associated antigen (ATLA) in sera include indirect immunofluorescence (7), particle-agglutination (8), ELISA (9), and Western-blot analysis (10). It has been argued that HTLV-I is potentially transmissible through do- nated blood and that transfusion of risky blood should be avoided (43). Human T-cell lymphotropic virus type 1 (HTLV-I) has been associated with adult T-cell leukemia (ATL) and HTLV-I-associated myelopathy (HAM) (1-3). enzvmatic polymerization, dot-blot assay, oligonucleotide probe, genomk structure Therefore, it can be considered to be a powerful analyt- ical system for the confirmation of HTLV-I infections. The PCR dot-blot assay system is simpleР вЂќР вЂћnd rapid for the detection of the target DNA sequence, although it does not pro- vide useful additional information, as does the Southern-blot analysis. Furthermore, all data ob- tained with the specimens tested were confirmed by the PCR oligomer-restriction (OR) procedure. Test and revealed faint banding patterns in Western-blot analysis, the PCR dot-blot as- say system can clearly detect the HTLV-I provirus DNA. Al- though carriers sometimes showed low an- tibody titers in the particle-agglutination We checked genomic DNAs extractedР вЂќР вЂЎrom peripheral blood lymphocytes (PBL) of five adult T-cell leukemia patients, one HTLV-I-associated myelopathy (HAM) pa- tient, four asymptomatic carriers, and five normal individuals as negative controls.

DNA sequences coding the p24 in the gag re- gion were chosen as targets to be amplified. Mitsutoshi Tara2 Diagnostics Division, Central Research Laboratories, Fujirebio, Inc., Tokyo and 2Department of Internal Medicine, Kagoshima City Hospital, Kagoshima, JapanР вЂќР вЂЎor the detection of the HTLV-I provirus genomic DNA, we applied the polymerase- chain-reaction (PCR) dot-blot assay sys- tem developed by Saiki et al. Journal of Clinical Laboratory Analysis 2:194-198 (1988)Р вЂќР’Вetection of Human T-cell Lymphotropic Virus Type-I Provirus DNA by In Vitro EnzymaticР вЂќР вЂћmplification Nobuyuki Fujii, Masanori Ogasawara, Satoru Ito, and

Dot blots image pro plus