Rapid,sensitive, specific,and quantitative detection of human T-cell leukemia virus type 1 sequence in peripheral blood mononuclear cells by an improved polymerase chain reaction method with nested primers |
| |
Authors: | Yuko Aono Joko Imai Keiko Tominaga Satoshi Orita Akihiko Sato Hisanaga Igarashi |
| |
Affiliation: | (1) Shionogi Institute for Medical Science, 566 Settsu-shi, Osaka, Japan;(2) Institute for Virus Research, Kyoto University, Kyoto, Japan |
| |
Abstract: | Improving on the nested double polymerase chain reaction (PCR) described previously, we have developed a new two-step PCR (TS-PCR) method for detecting more specifically the human T-cell leukemia virus type 1 (HTLV-1) proviral sequences in peripheral blood mononuclear cells (PBMC). In our TS-PCR method, the point of modification is to use optimal concentrations of primers in the first amplification step in the range of 0.01–0.025 µM. This increases sensitivity and specificity enough to detect from 1 to 105 copies of template DNA without radioisotopes. This method is rapid because of completion in 1 day and is also applicable for quantitative detection of clinical specimens. The data show that the quantitative detection of HTLV-1 proviral sequences by this method correlates with the anti-HTLV-1 antibody titers from serologic analysis of seropositive healthy carriers. Moreover, the TS-PCR method using each specific primer was also attempted for successful detection of other viral genomes; therefore, the principle of this method is widely suitable for routine detection of genomes in the basic and clinical microbiological fields. |
| |
Keywords: | human T-cell leukemia virus type 1 HTLV-1 polymerase chain reaction nested primers HTLV-1 healthy carriers |
本文献已被 SpringerLink 等数据库收录! |
|