Semiquantitative analysis of integrated genomes of human T-lymphotropic virus type I in asymptomatic virus carriers.
Semiquantitative analysis of integrated genomes of human T-lymphotropic virus type I in asymptomatic virus carriers.
复制标题
无症状病毒携带者中人类 T 淋巴细胞病毒 I 型整合基因组的半定量分析。
作者：
O. Shinzato;S. Ikeda;S. Momita;Y. Nagata;S. Kamihira;E. Nakayama;H. Shiku
半定量估计人T淋巴细胞病毒I型（HTLV-I）的整合外周血单核细胞（PBMC）进行。对来自134个HTLV-1携带者的基因组DNA样品进行40或60个循环的聚合酶链反应以扩增HTLV-1的pol区域。用32 P标记的寡核苷酸探针对pol区进行斑点杂交，检测HTLV-Ⅰ基因组。然后用RI成像系统（Ambis Inc，San Diego，CA）计数杂交点膜的放射性，并通过与连续稀释的HTLV-I基因组阳性DNA的标准曲线比较来确定HTLV-I基因组剂量。观察到HTLV-I基因组整合的广泛变化。当整合基因组剂量计算为每100个PBMC中HTLV-I拷贝数时，7名携带者（5%）具有超过10个拷贝，56名（42%）具有1至10个拷贝，46名（34%）具有0.1至1个拷贝，24名（18%）具有小于0.1个拷贝。在一个样品中，HTLV-I基因组是不可检测的，这可能表明整合的基因组以每100个PBMC小于0.01个拷贝存在。不同HTLV-I基因组个体分布的年龄或性别相关变化相当有限。然而，HTLV-I基因组水平高的携带者总是超过30岁，并且主要是男性（7人中有6人）。
A semiquantitative estimation of human T-lymphotropic virus type I (HTLV-I) integration by peripheral blood mononuclear cells (PBMC) was performed. Genomic DNA samples derived from 134 HTLV-I carriers were subjected to 40 or 60 cycles of the polymerase chain reaction to amplify the pol region of HTLV-I. The HTLV-I genome was detected by dot hybridization using a 32P-labeled oligonucleotide probe for the pol region. The radioactivity of hybridized dot membranes was then counted with an RI Imaging System (Ambis Inc, San Diego, CA) and the HTLV-I genome dose was determined by comparison with standard curve for serially diluted HTLV-I genome-positive DNA. A wide range of variation of HTLV-I genome integration was observed. When the integrated genome dose was calculated as the number of HTLV-I copies per 100 PBMC, 7 carriers (5%) had more than 10 copies, 56 (42%) had 1 to 10 copies, 46 (34%) had 0.1 to 1 copy, and 24 (18%) had less than 0.1 copy. In one sample, the HTLV-I genome was undetectable, which may indicate that the integrated genome was present at less than 0.01 copies per 100 PBMC. Age- or sex-related variations in the distribution of individuals with different HTLV-I genome were rather limited. However, carriers with a high level of the HTLV-I genome were always more than 30 years old and were predominantly male (six of seven).