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

Overview

Journal Virus Genes

Specialties Microbiology
Molecular Biology

Date 1992 Apr 1

PMID 1589963

Citations 4

Authors

Y Aono

J Imai

K Tominaga

S Orita

A Sato

H Igarashi

Affiliations

Soon will be listed here.

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 microM. This increases sensitivity and specificity enough to detect from 1 to 10(5) 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.

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