Tuke P W, Luton P, Garson J A
Department of Medical Microbiology, University College and Middlesex School of Medicine, London, UK.
J Virol Methods. 1992 Nov;40(2):163-73. doi: 10.1016/0166-0934(92)90065-l.
A 'nested' polymerase chain reaction (PCR) assay is described which is capable of detecting single copies of human T-cell lymphotropic virus (HTLV) in genomic DNA extracted from peripheral blood mononuclear cells (PBMCs). A single set of 'nested' oligonucleotide primers, based on the highly conserved tax/rex region of the viral genome, was able to detect both HTLV-I and HTLV-II proviral sequences in clinical samples of diverse geographical origins, from the United States, Great Britain, Japan, the Caribbean, Italy, Greece, Iraq and West Africa. Rapid discrimination between HTLV-I and HTLV-II infections was achieved by restriction enzyme analysis of unpurified second-round PCR products, even in those cases in which serological assays had failed to provide a definitive result. Over a 2-year period, a total of 53 HTLV infections (37 HTLV-I and 16 HTLV-II) were identified by this technique and complete concordance with serological typing, available in 41 cases, was observed.
本文描述了一种“巢式”聚合酶链反应（PCR）检测方法，该方法能够在外周血单个核细胞（PBMC）提取的基因组DNA中检测到单拷贝的人类嗜T淋巴细胞病毒（HTLV）。基于病毒基因组高度保守的tax/rex区域设计的一组“巢式”寡核苷酸引物，能够在来自美国、英国、日本、加勒比地区、意大利、希腊、伊拉克和西非等不同地理区域的临床样本中检测到HTLV - I和HTLV - II前病毒序列。通过对未纯化的第二轮PCR产物进行限制性酶切分析，即使在血清学检测未能给出明确结果的情况下，也能快速区分HTLV - I和HTLV - II感染。在两年的时间里，通过该技术共鉴定出53例HTLV感染（37例HTLV - I和16例HTLV - II），并且在41例可获得血清学分型结果的病例中，观察到与血清学分型完全一致。
