A novel single cell PCR assay: detection of human T lymphotropic virus type I DNA in lymphocytes of patients with adult T cell leukemia.
The abnormal lymphocytes in adult T cell leukemia (ATL) reveal a peculiar morphology that is characterized by indented or lobulated nuclei. While human T lymphotropic virus type I (HTLV-I) is thought to be integrated in ATL cells, the correlation between the nuclear irregularities and HTLV-I infection is obscure. We have devised a novel single cell polymerase chain reaction (PCR) technique to examine the integration of HTLV-I provirus genome in cells from two patients with ATL. To isolate single cells, peripheral blood smears were prepared on thin polyester slides and stained with May-Grünwald-Giemsa. Morphologically defined single cells were cut out after light microscopy. The HTLV-I DNA sequences were detected not only in ATL cells but also in normal-looking lymphocytes. This novel PCR method may provide a valuable tool for understanding the molecular events associated with HTLV-I infection at the single cell level.
Miyagi
T
T
Department of Medicine, Kochi Medical School, Japan.
Murakami
K
K
Sawada
T
T
Taguchi
H
H
Miyoshi
I
I
D000328:Adult; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D015368:Human T-lymphotropic virus 1 / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D015459:Leukemia-Lymphoma, Adult T-Cell / Q000473:pathology / Q000821:virology*; D008214:Lymphocytes / Q000473:pathology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D004279:DNA, Viral