دورية أكاديمية
Rapid and Sensitive Qualitative Duoplex Real-Time PCR Method for Discriminatory and Confirmatory Diagnosis of HTLV-1 and HTLV-2 Infections: Brazilian Multicentric Study.
| العنوان: | Rapid and Sensitive Qualitative Duoplex Real-Time PCR Method for Discriminatory and Confirmatory Diagnosis of HTLV-1 and HTLV-2 Infections: Brazilian Multicentric Study. |
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| المؤلفون: | Rocha-Junior MC; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Graduate Program in Biosciences and Biotechnology, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, São Paulo, Brazil., Rodrigues ES; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Center for Cell-Based Therapy CTC, Regional Blood Center of Ribeirão Preto, University of São Paulo, São Paulo, Brazil., Slavov SN; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Center for Cell-Based Therapy CTC, Regional Blood Center of Ribeirão Preto, University of São Paulo, São Paulo, Brazil., Assone T; Laboratory of Medical Investigation LIM 56, Division of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil., Pedreschi M; Laboratory of Medical Investigation LIM 56, Division of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil., de La Roque DGL; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Tropical Medicine Center, Federal University of Para, Belém, Brazil., Sousa M; Tropical Medicine Center, Federal University of Para, Belém, Brazil., Olavarria V; Centro de HTLV, Escola Bahiana de Medicina e Saúde Pública, Salvador, Brazil., Galvão-Castro B; Centro de HTLV, Escola Bahiana de Medicina e Saúde Pública, Salvador, Brazil., da Fonseca BAL; Department of Neurology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil., Penalva de Oliveira AC; Laboratory of Medical Investigation LIM 56, Division of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil., Smid J; Laboratory of Medical Investigation LIM 56, Division of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil., Takayanagui OM; Department of Neurology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil., Casseb J; Laboratory of Medical Investigation LIM 56, Division of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil., Covas DT; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Center for Cell-Based Therapy CTC, Regional Blood Center of Ribeirão Preto, University of São Paulo, São Paulo, Brazil.; Department of Neurology, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil., Kashima S; Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical School, University of São Paulo, São Paulo, Brazil.; Graduate Program in Biosciences and Biotechnology, School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, São Paulo, Brazil.; Center for Cell-Based Therapy CTC, Regional Blood Center of Ribeirão Preto, University of São Paulo, São Paulo, Brazil. |
| المصدر: | Frontiers in medicine [Front Med (Lausanne)] 2022 Jun 09; Vol. 9, pp. 881630. Date of Electronic Publication: 2022 Jun 09 (Print Publication: 2022). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Frontiers Media S.A Country of Publication: Switzerland NLM ID: 101648047 Publication Model: eCollection Cited Medium: Print ISSN: 2296-858X (Print) Linking ISSN: 2296858X NLM ISO Abbreviation: Front Med (Lausanne) Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Lausanne, Switzerland : Frontiers Media S.A., [2014]- |
| مستخلص: | Human T cell lymphotropic virus (HTLV) is the caustive agent of two main conditions i. e., the HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) and the adult T-cell leukemia/lymphoma (ATLL). HTLV diagnosis is based on serological and molecular approaches; however, an accurate and validated method is still needed. The objective of this study was to establish a rapid and sensitive molecular test to confirm and discriminate HTLV 1/2 types. The test validation was performed as a multicentric study involving HTLV confirmation centers throughout Brazil. Proviral DNA was extracted from whole blood and the amplification was performed using in-house designed primer and probe sets targeting the pol genomic region. An internal control to validate the extraction and amplification was also included. The limit of detection (LoD) of the assay was four copies/reaction for HTLV-1 and 10.9 copies/reaction for HTLV-2. The diagnostic sensitivity of the platform was 94.6% for HTLV-1, 78.6% for HTLV-2, and the specificity was 100% for both viruses. Cross-reactions of the test with human viruses including HAV, HBV, HCV, HIV-1/2, and parvovirus B19 were not observed. During the multicentric validation, the test was used to screen a total of 692 blood samples obtained from previously confirmed HTLV-positive individuals. From these, 91.1% tested positive being concordant with the previously obtained results. In conclusion, our duoplex-RT-PCR-HTLV1 /2 presented adequate efficiency for HTLV-1/2 differentiation showing high sensitivity and specificity. Therefore, it can be a suitable tool for confirmation of suspected and inconclusive HTLV cases, prenatal and pre-transplant diagnosis, in Brazil and in other countries HTLV-endemic countries. Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2022 Rocha-Junior, Rodrigues, Slavov, Assone, Pedreschi, de La Roque, Sousa, Olavarria, Galvão-Castro, Fonseca, Penalva de Oliveira, Smid, Takayanagui, Casseb, Covas and Kashima.) |
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| فهرسة مساهمة: | Keywords: HTLV-1; HTLV-2; molecular diagnosis; multiplex real time PCR; real time PCR |
| تواريخ الأحداث: | Date Created: 20220627 Latest Revision: 20220716 |
| رمز التحديث: | 20221213 |
| مُعرف محوري في PubMed: | PMC9218175 |
| DOI: | 10.3389/fmed.2022.881630 |
| PMID: | 35755037 |
| قاعدة البيانات: | MEDLINE |
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