Real-time polymerase chain reaction for quantification of HTLV-1 proviral load: application for analyzing aberrant integration of the proviral DNA in adult T-cell leukemia.
We describe the establishment of a real-time polymerase chain reaction (PCR) assay for the quantitative estimation of human T-cell leukemia virus type 1 (HTLV-1) proviral load using a LightCycler Technology (Roche Diagnostics, Mannheim, Germany) instrument. Proviral DNA level represents a measure of the integrated viral genome in host cells, so we applied this technique to evaluate the tumor burden in adult T-cell leukemia (ATL) patients with aberrant integration patterns of HTLV-1 detected by standard Southern blot hybridization (SBH) analysis. In 14 of our 15 ATL cases with 2 or more bands detected by SBH analysis, the ATL cells were shown to harbor multiple copies of the provirus within 1 ATL cell. This result suggests the usefulness of real-time PCR quantification for the study of the relationship between ATL pathology and HTLV-1-induced pathogenesis.
Kamihira
S
S
Department of Laboratory Medicine, Nagasaki University School of Medicine, Japan. kamihira@net.nagasaki-u.ac.jp
Dateki
N
N
Sugahara
K
K
Yamada
Y
Y
Tomonaga
M
M
Maeda
T
T
Tahara
M
M
D004279:DNA, Viral / Q000097:blood; D018628:Gene Dosage; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000254:growth & development; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D011533:Proviruses / Q000254:growth & development*; D012015:Reference Standards; D019562:Viral Load / Q000379:methods* / Q000592:standards; D016662:Virus Integration
D004279:DNA, Viral