1. Low-cost and simple PCR process for access to molecular diagnosis of HTLV-1/2 in low-resource countries.
- Author
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Ducasa N, Domínguez D, Benencio P, Alfie L, Etcheves P, Scarton G, Biglione M, and Caputo M
- Subjects
- Humans, Retrospective Studies, Real-Time Polymerase Chain Reaction methods, Real-Time Polymerase Chain Reaction economics, Developing Countries, Polymerase Chain Reaction methods, Polymerase Chain Reaction economics, Human T-lymphotropic virus 1 genetics, Human T-lymphotropic virus 1 isolation & purification, Human T-lymphotropic virus 2 genetics, Human T-lymphotropic virus 2 isolation & purification, HTLV-I Infections diagnosis, HTLV-I Infections virology, Sensitivity and Specificity, HTLV-II Infections diagnosis, Molecular Diagnostic Techniques methods, Molecular Diagnostic Techniques economics
- Abstract
Background: HTLV-1/2 exhibit a widespread distribution globally and are associated with severe clinical manifestations, necessitating precise viral identification for diagnosis. Currently, there are no official diagnostic guidelines, and a variety of published protocols exists. We introduce an enhanced nested real-time PCR technique followed by high-resolution melting (rtPCR-HRM), designed to offer a cost-effective and straightforward tool for the simultaneous identification of both viruses., Methods: The technique was tested in a retrospective, blinded study, involving a total panel of 110 samples, of which 47 were positive for HTLV-1, 12 for HTLV-2, and 51 tested negatives. Additionally, we compared the performance of this technique with a line immunoassay (LIA)., Results: The results demonstrate 100 % sensitivity, specificity, and diagnostic accuracy for both viruses. Sensitivity analysis indicated that at least 1 viral copy of HTLV-1 and 14.4 viral copies of HTLV-2 could be reliably detected., Conclusions: Our results indicate that rtPCR-HRM is effective in confirming HTLV-1 and HTLV-2 infection, important in Latin American countries where both viruses circulate. Furthermore, the proposed strategy provides a new tool that can be used to resolve indeterminate cases identified by Western blot, with the added advantage of being faster and simpler than n-PCR and more cost-effective than other probe-based RT-PCRs., Competing Interests: Declaration of competing interest No conflict of interest declared., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
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