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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art_ROCHA-JUNIOR_Rapid_and_Sensitive_Qualitative_Duoplex_RealTime_PCR_Method_2022.PDF (1.15 MB)
Citações na Scopus
2
Tipo de produção
article
Data de publicação
2022
DOI
Scopus
Web of Science
PubMed
Título da Revista
ISSN da Revista
Título do Volume
Editora
FRONTIERS MEDIA SA
Autores
ROCHA-JUNIOR, Mauricio Cristiano
RODRIGUES, Evandra Strazza
SLAVOV, Svetoslav Nanev
ROQUE, Debora Glenda Lima de la
SOUSA, Maisa
OLAVARRIA, Viviana
GALVAO-CASTRO, Bernardo
FONSECA, Benedito Antonio Lopes da
Citação
FRONTIERS IN MEDICINE, v.9, article ID 881630, 9p, 2022
Projetos de Pesquisa
Unidades Organizacionais
Fascículo
Resumo
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.
Palavras-chave
HTLV-1, HTLV-2, molecular diagnosis, real time PCR, multiplex real time PCR
URI
https://observatorio.fm.usp.br/handle/OPI/48371
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Coleções
Artigos e Materiais de Revistas Científicas - FM/MDT
Artigos e Materiais de Revistas Científicas - HC/ICHC
Artigos e Materiais de Revistas Científicas - HC/IMREA
Artigos e Materiais de Revistas Científicas - LIM/56
Artigos e Materiais de Revistas Científicas - ODS/03