A new diagnostic algorithm using biopsy specimens in adult T-cell leukemia/lymphoma: combination of RNA in situ hybridization and quantitative PCR for HTLV-1.
A new diagnostic algorithm using biopsy specimens in adult T-cell leukemia/lymphoma: combination of RNA in situ hybridization and quantitative PCR for HTLV-1.
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DOI：
10.1038/s41379-020-0635-8
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Karube K
中科院分区：
文献类型：
--
作者：
Takatori M;Sakihama S;Miyara M;Imaizumi N;Miyagi T;Ohshiro K;Nakazato I;Hayashi M;Todoroki J;Morishima S;Masuzaki H;Fukushima T;Karube K
成人t细胞白血病/淋巴瘤（ATLL）和其他t细胞肿瘤之间的组织病理学区别往往具有挑战性。目前准确诊断ATLL的金标准是Southern blot杂交（SBH）试验，该试验检测人t细胞白血病病毒I型（HTLV-1）前病毒的克隆整合。然而，SBH不能用小活检或福尔马林固定石蜡包埋（FFPE）组织样本进行，因为这种检测需要大量未降解的DNA。在这里，我们开发了一种新的诊断算法，用于使用FFPE样本准确诊断ATLL。该方法结合了HTLV-1的两种检测方法，即利用RNAscope进行HTLV-1 bZIP因子的超灵敏RNA原位杂交（HBZ-RNAscope）和针对tax基因的定量PCR （tax- qpcr）。我们分析了119个FFPE组织样本（62个ATLL， 57个非ATLL，包括41个HTLV-1携带者），并将其与使用相应新鲜冷冻样本的SBH结果进行了比较。因此，tax-qPCR的ATLL识别率高于HBZ-RNAscope（分别为88%[52/59]和63%[39/62]）。然而，HBZ-RNAscope清晰地显示了htlv -1感染肿瘤细胞的定位，当分析仅限于2岁以下的样本时，其识别率提高到94%(17/18)，表明其在日常诊断中的实用性。结合这两种检测方法的诊断算法成功评估了94%（112/119）的样本，并以100%的灵敏度和特异性将ATLL与包括HTLV-1携带者在内的非ATLL病例区分开来。该方法有望取代SBH，提高ATLL的诊断准确性。
Histopathological distinction between adult T-cell leukemia/lymphoma (ATLL) and other T-cell neoplasms is often challenging. The current gold standard for the accurate diagnosis of ATLL is the Southern blot hybridization (SBH) assay, which detects clonal integration of human T-cell leukemia virus type I (HTLV-1) provirus. However, SBH cannot be performed with small biopsy or formalin-fixed paraffin-embedded (FFPE) tissue samples because this assay requires a large amount of DNA without degradation. Here we developed a new diagnostic algorithm for the accurate diagnosis of ATLL using FFPE samples. This method combines two HTLV-1 detection assays, namely, ultrasensitive RNA in situ hybridization using RNAscope for HTLV-1 bZIP factor (HBZ-RNAscope), and quantitative PCR targeting the tax gene (tax-qPCR). We analyzed 119 FFPE tissue specimens (62 ATLL, and 57 non-ATLL, including 41 HTLV-1 carriers) and compared them with the SBH results using the corresponding fresh-frozen samples. As a result, tax-qPCR had a higher ATLL identification rate than HBZ-RNAscope (88% [52/59], and 63% [39/62], respectively). However, HBZ-RNAscope clearly visualized the localization of HTLV-1-infected tumor cells and its identification rate increased to 94% (17/18) when the analysis was limited to samples up to 2 years old, indicating its usefulness in the daily diagnosis. The diagnostic algorithm combining these two assays successfully evaluated 94% (112/119) of samples and distinguished ATLL from non-ATLL cases including HTLV-1 carriers with 100% sensitivity and specificity. This method is expected to replace SBH and increase the accuracy of the diagnosis of ATLL.
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DOI：
10.3390/v7122952
发表时间：
2015-12-09
期刊：
Viruses
影响因子：
--
作者：
Mesnard JM;Barbeau B;Césaire R;Péloponèse JM
通讯作者：
Péloponèse JM
DOI：
10.1073/pnas.78.10.6476
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
HINUMA, Y;NAGATA, K;MIYOSHI, I
通讯作者：
MIYOSHI, I
影响因子：
4.5
作者：
Kaneshiro, Yukako;Sourinphoumy, Khamphang;Fukushima, Takuya
通讯作者：
Fukushima, Takuya
影响因子：
5.6
作者：
Karube, Kennosuke;Suzumiya, Junji;Ohshima, Koichi
通讯作者：
Ohshima, Koichi
DOI：
10.1073/pnas.0507631103
发表时间：
2006-01-17
影响因子：
11.1
作者：
Satou, Y;Yasunaga, J;Matsuoka, M
通讯作者：
Matsuoka, M