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J Appl Microbiol ; 129(3): 768-774, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32202037

RESUMEN

AIMS: To evaluate the potential use of synthetic oligonucleotides as a standard curve for proviral load (PVL) of human T-cell leukaemia virus type 1 (HTLV-1) quantification in peripheral blood mononuclear cells (PBMC) of HTLV-1-infected individuals by quantitative real-time polymerase chain reaction (qPCR) analysis. METHODS AND RESULTS: Synthetic oligonucleotides based on HTLV-1 genome were customized to use as a standard curve. Twelve anti-HTLV-1-positive samples with known HTLV-1 PVL, previously quantified by qPCR assay using TARL-2 cells as a conventional standard curve, were submitted to the new protocol. The proviral quantification levels had a high concordance with qPCR results using a conventional standard curve. The results demonstrate that the conventional standard curve can be replaced by a synthetic standard curve due to its ability to quantification based on the linearity and qPCR efficiency and similar results with a validated qPCR assay using a conventional standard curve. CONCLUSIONS: Synthetic oligonucleotides standard curves could be a very useful tool on HTLV-1 diagnosis and absolute HTLV-1 PVL quantification. SIGNIFICANCE AND IMPACT OF THE STUDY: HTLV-1 PVL determination using synthetic oligonucleotides standard curve by qPCR could be a helpful alternative for the laboratories that monitor infected patients as an important prognostic factor in HTLV-1-associated diseases progression. Also, it can decrease costs and overcome the biological limitations of the plasmid curve.


Asunto(s)
Infecciones por HTLV-I/diagnóstico , Virus Linfotrópico T Tipo 1 Humano/aislamiento & purificación , Provirus/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Carga Viral/métodos , Adulto , ADN Viral/genética , Progresión de la Enfermedad , Genoma Viral/genética , Infecciones por HTLV-I/sangre , Infecciones por HTLV-I/virología , Virus Linfotrópico T Tipo 1 Humano/genética , Humanos , Leucocitos Mononucleares/virología , Persona de Mediana Edad , Oligonucleótidos/síntesis química , Oligonucleótidos/genética , Pronóstico , Provirus/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/normas , Carga Viral/normas
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