Cardoso E A, Miranda N, Gameiro P, Frade M J, Figueiredo M, Parreira A
Laboratório de Biologia Molecular (CIPM), Instituto Portugués de Oncologia, Lisboa, Portugal.
J Clin Pathol. 1996 Nov;49(11):938-41. doi: 10.1136/jcp.49.11.938.
To study the expression of the human T lymphotropic virus (HTLV) Tax gene in peripheral blood mononuclear cells.
Blood was collected from 72 patients with lymphoproliferative disorders. Serum from all patients was assayed for antibodies directed against HTLV-I structural proteins by ELISA and western blotting. RNA was purified from fresh blood cells and amplified by reverse transcription polymerase chain reaction (RT-PCR). After Southern blotting, the PCR products were hybridised with a 32P end-labelled probe specific for the Tax gene.
All samples were seronegative. A specific band for the Tax gene was found in five samples. Each of the patients positive for Tax gene expression had a different type of lymphoproliferative disorder.
Infection by HTLV-I cannot be assessed solely by immunological assays, particularly when only disrupted virions are used. Sensitive molecular biology assays are essential for detecting viral gene expression in fresh blood cells.
研究人类嗜T淋巴细胞病毒(HTLV)Tax基因在外周血单个核细胞中的表达。
采集72例淋巴增殖性疾病患者的血液。采用酶联免疫吸附测定(ELISA)和蛋白质印迹法检测所有患者血清中针对HTLV-I结构蛋白的抗体。从新鲜血细胞中纯化RNA,并通过逆转录聚合酶链反应(RT-PCR)进行扩增。经过Southern印迹后,将PCR产物与Tax基因特异性的32P末端标记探针杂交。
所有样本血清学均为阴性。在5个样本中发现了Tax基因的特异性条带。Tax基因表达呈阳性的每位患者均患有不同类型的淋巴增殖性疾病。
不能仅通过免疫学检测来评估HTLV-I感染,特别是当仅使用破碎的病毒体时。灵敏的分子生物学检测对于检测新鲜血细胞中的病毒基因表达至关重要。