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 105 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.

中文翻译：

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人类T细胞白血病病毒1和-2多重定量PCR的开发和评估。

在日本，通常通过血清学检查来诊断人类1型T细胞白血病病毒（HTLV-1）感染，但Western印迹结果不确定性很高，因此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感染HTLV-1和/或HTLV-2，也用于HTLV-1相关疾病的随访。为了制备能够区分HTLV-1和HTLV-2感染的高灵敏度方法，开发了通过大规模引物筛选的多重定量聚合酶链反应（qPCR）。通过连续稀释细胞系并用已知的临床样品进行测试来评估敏感性和特异性。产生的多重qPCR可以每105个细胞检测到大约四个拷贝的HTLV-1前病毒。此外，在HTLV-1血清阳性临床样本中有97.2％（在211个样本中有205个）中检测到HTLV-1前病毒。与以前报道的方法相比，这些灵敏度足够高。同样，通过该方法发现所有测试的HTLV-2血清反应阳性临床样品均为阳性（三分之三）。综上所述，该方法能够以极高的灵敏度成功并同时检测HTLV-1和HTLV-2前病毒。