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 posi...