Detection of HTLV-I pX gene by polymerase chain reaction using newly designed primers.

Abstract

Newly designed oligonucleotide primers, KI-7 and KI-8 for the human T cell lymphotropic virus type I (HTLV-I) pX gene were synthesized using an automated DNA synthesizer. Previously known HTLV-I-infected cell lines, MT-1 and MT-2, were used as positive controls and HTLV-I-uninfected cell lines, Molt-4, SBC-3, ABC-1, and EBC-1, as negative controls. Peripheral blood mononuclear cells from 17 patients with anti-HTLV-I antibody and 10 healthy individuals without anti-HTLV-I antibody were studied by polymerase chain reaction (PCR) with KI-7 and KI-8. All DNA samples from HTLV-I-infected cell lines and 17 patients with anti-HTLV-I antibodies showed positive signals of the HTLV-I pX gene. None of the DNA samples from HTLV-I-uninfected cell lines or 10 healthy individuals showed positive signals. When serially diluted DNA of MT-2 cells were amplified by 35 cycles of PCR, the detection limit of the pX gene by using the primer pairs was DNA from about 1.5 MT-2 cells. Specificity and detectable capacity of primer pairs, KI-7 and KI-8 were confirmed to be enough to use for the diagnosis of HTLV-I infection.

• Authors: K Imajo, K Shinagawa, S Tada, T Tsubota, I Kimura
• Journal: Acta medica Okayama (Acta Med Okayama) Vol. 47 Issue 6 Pg. 355-61 (Dec 1993) ISSN: 0386-300X [Print] Japan
• PMID: 8128908 (Publication Type: Journal Article)

Chemical References

• DNA, Viral
• HTLV-I Antibodies
• Oligonucleotide Probes

Topics

• Base Sequence
• Blotting, Southern
• Cell Line
• DNA, Viral (analysis)
• Genes, pX
• HTLV-I Antibodies (analysis)
• Human T-lymphotropic virus 1 (genetics)
• Humans
• Molecular Sequence Data
• Oligonucleotide Probes (genetics)
• Polymerase Chain Reaction (methods)