Department of Safety Research on Blood and Biological Products, National Institute of Infectious Diseases, Tokyo, Japan.
Department of Quality, Japanese Red Cross Kyushu Block Blood Center, Fukuoka, Japan.
Microbiol Immunol. 2019 Nov;63(11):458-464. doi: 10.1111/1348-0421.12740. Epub 2019 Sep 11.
The diagnosis of human T -cell leukemia virus type 1 (HTLV-1) infection in Japan is usually performed by serological testing, but the high rate of indeterminate results from western blotting makes it difficult to assess the infection accurately. Nucleic acid tests for HTLV-1 and/or HTLV-2 are used to confirm infection with HTLV-1 and/or HTLV-2 and are also used for the follow-up of HTLV-1 related diseases. To prepare a highly sensitive method that can discern infection with HTLV-1 and HTLV-2, a multiplex quantitative polymerase chain reaction (qPCR) by large-scale primer screening was developed. Sensitivity and specificity were evaluated by serial dilution of cell lines and by testing with known clinical samples. The resulting multiplex qPCR can detect about four copies of HTLV-1 provirus per 10 cells. Moreover, HTLV-1 provirus could be detected in 97.2% (205 of 211) of HTLV-1 seropositive clinical samples. These sensitivities were sufficiently high compared with the methods reported previously. Also, all the HTLV-2 seropositive clinical samples tested were found to be positive by this method (three of three). In conclusion, this method can successfully and simultaneously detect both types of HTLV-1 and HTLV-2 provirus with extremely high sensitivity.
在日本，人类 T 细胞白血病病毒 1 型（HTLV-1）感染的诊断通常通过血清学检测进行，但 Western blot 的不确定结果率较高，使得难以准确评估感染情况。HTLV-1 和/或 HTLV-2 的核酸检测用于确认 HTLV-1 和/或 HTLV-2 的感染，也用于 HTLV-1 相关疾病的随访。为了制备一种能够区分 HTLV-1 和 HTLV-2 感染的高度敏感方法，通过大规模引物筛选开发了一种多重定量聚合酶链反应（qPCR）。通过细胞系的连续稀释和已知临床样本的检测来评估灵敏度和特异性。该多重 qPCR 可检测到约每 10 个细胞中有 4 个拷贝的 HTLV-1 前病毒。此外，该方法可检测到 97.2%（205/211）的 HTLV-1 血清阳性临床样本中的 HTLV-1 前病毒。与之前报道的方法相比，这些灵敏度足够高。此外，该方法还可检测到所有 3 份 HTLV-2 血清阳性的临床样本均为阳性。总之，该方法可以成功且同时以极高的灵敏度检测两种类型的 HTLV-1 和 HTLV-2 前病毒。
