Kato Junki, Masaki Ayako, Fujii Keiichiro, Takino Hisashi, Murase Takayuki, Yonekura Kentaro, Utsunomiya Atae, Ishida Takashi, Iida Shinsuke, Inagaki Hiroshi
Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Department of Dermatology, Imamura Bun-in Hospital, Kagoshima, Japan.
Pathol Int. 2016 Nov;66(11):618-621. doi: 10.1111/pin.12462. Epub 2016 Sep 27.
Detection of HTLV-1 provirus using paraffin tumor sections may assist the diagnosis of adult T-cell leukemia/lymphoma (ATLL). For the detection, non-quantitative PCR assay has been reported, but its usefulness and limitations remain unclear. To our knowledge, quantitative PCR assay using paraffin tumor sections has not been reported. Using paraffin sections from ATLLs and non-ATLL T-cell lymphomas, we first performed non-quantitative PCR for HTLV-1 provirus. Next, we determined tumor ratios and carried out quantitative PCR to obtain provirus copy numbers. The results were analyzed with a simple regression model and a novel criterion, cut-off using 95 % rejection limits. Our quantitative PCR assay showed an excellent association between tumor ratios and the copy numbers (r = 0.89, P < 0.0001). The 95 % rejection limits provided a statistical basis for the range for the determination of HTLV-1 involvement. Its application suggested that results of non-quantitative PCR assay should be interpreted very carefully and that our quantitative PCR assay is useful to estimate the status of HTLV-1 involvement in the tumor cases. In conclusion, our quantitative PCR assay using paraffin tumor sections may be useful for the screening of ATLL cases, especially in HTLV-1 non-endemic areas where easy access to serological testing for HTLV-1 infection is limited.
使用石蜡包埋的肿瘤切片检测人嗜T淋巴细胞病毒1型（HTLV-1）前病毒可能有助于成人T细胞白血病/淋巴瘤（ATLL）的诊断。对于该检测，已有非定量PCR检测方法的报道，但其有效性和局限性仍不明确。据我们所知，尚未有使用石蜡包埋肿瘤切片进行定量PCR检测的报道。我们使用ATLL和非ATLL T细胞淋巴瘤的石蜡切片，首先对HTLV-1前病毒进行非定量PCR检测。接下来，我们确定肿瘤比例并进行定量PCR以获得前病毒拷贝数。结果采用简单回归模型和一种新的标准（使用95%排除限的截断值）进行分析。我们的定量PCR检测显示肿瘤比例与拷贝数之间具有良好的相关性（r = 0.89，P < 0.0001）。95%排除限为确定HTLV-1感染情况提供了统计学依据。其应用表明，非定量PCR检测结果的解读应非常谨慎，并且我们的定量PCR检测对于评估肿瘤病例中HTLV-1感染状态是有用的。总之，我们使用石蜡包埋肿瘤切片的定量PCR检测可能有助于ATLL病例的筛查，尤其是在难以进行HTLV-1感染血清学检测的HTLV-1非流行地区。
