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
| Autor: | Shugo Sakihama, Kazuiku Ohshiro, Satoko Morishima, Mitsuyoshi Takatori, Megumi Miyara, Takashi Miyagi, Kennosuke Karube, Takuya Fukushima, Junpei Todoroki, Hiroaki Masuzaki, Masaki Hayashi, Iwao Nakazato, Naoki Imaizumi |
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| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Pathology medicine.medical_specialty Biopsy In situ hybridization Biology Real-Time Polymerase Chain Reaction Sensitivity and Specificity Article Adult T-cell leukemia/lymphoma Pathology and Forensic Medicine 03 medical and health sciences 0302 clinical medicine immune system diseases hemic and lymphatic diseases medicine Humans Leukemia-Lymphoma Adult T-Cell Tumour virus infections In Situ Hybridization Southern blot Human T-lymphotropic virus 1 Deltaretrovirus Infections medicine.diagnostic_test Gold standard (test) Provirus medicine.disease Leukemia 030104 developmental biology Real-time polymerase chain reaction 030220 oncology & carcinogenesis RNA Viral T-cell lymphoma Algorithm Algorithms |
| Zdroj: | Modern Pathology |
| ISSN: | 0893-3952 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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