Estandarización y validación de la técnica de Reacción en Cadena de la Polimerasa Nested Múltiple para el diagnóstico de HTLV-1

• Autor: SERRANO SEGURA, Kevin Osnar
• Formato: tesis de grado
• Fecha de Publicación: 2019
• Institución: Universidad Nacional de Trujillo
• Repositorio: UNITRU-Tesis
• Lenguaje: español
• OAI Identifier: oai:dspace.unitru.edu.pe:20.500.14414/12575
• Enlace del recurso: https://hdl.handle.net/20.500.14414/12575
• Nivel de acceso: acceso abierto
• Materia: HTLV-1; PCR nested; sensibilidad

In Peru, the existence of a normative diagnostic algorithm is unknown, it is assumed that it is similar to HIV, therefore a multiple nested PCR test was standardized and validated for the diagnosis of HTLV-1 in proviral DNA samples of reference material of the MT-2 cell line. The test was validated with proviral DNA samples from whole blood, it was determined in 70 samples characterized by serology, its sensitivity and analytical specificity, veracity and accuracy in comparison with the immunoblot test (gold standard). The results of the validation showed a limit of detection of 0.5 ng of DNA per reaction of 25 μL of the Pol and LTR gene and an accuracy with a 100% detection rate for 0.4, 0.5 and 0.6 ng of DNA. In addition, in comparison with an Immunoblot test (INNOLIA) considered a reference test for our study, the multiple nested PCR obtained 97% sensitivity, 100% specificity, a positive and negative predictive value of 97.5 and 99.9% respectively and a kappa match factor of 0.97. The standardized method (multiple nested PCR) in this study is suitable and valid for the diagnosis of HTLV-1 in whole blood samples and can be implemented in the diagnosis algorithm of HTLV in the National Reference Laboratory of VTS-HIV / AIDS and to resolve the indeterminate results for the confirmatory diagnosis of children under 18 months with a high sensitivity and specificity._x000D_ Key words: HTLV-1, nested PCR, sensitivity, specificity