Evaluación de la capacidad que tienen los niveles de ácido desoxirribonucleico libre circulante mitocondrial y nuclear urinario para discriminar muestras de sujetos con nefritis lúpica
Cargando...
Fecha
2022
Autores
Márquez Escobar, Olga Cecilia
Título de la revista
ISSN de la revista
Título del volumen
Editor
Ediciones Universidad Simón Bolívar
Facultad de Ciencias Básicas y Biomédicas
Facultad de Ciencias Básicas y Biomédicas
Resumen
El lupus eritematoso sistémico (LES), es una enfermedad autoinmune, proclive a una presentación clínica heterogénea que debuta de manera silente en casos de nefritis lúpica activa (NLA), lo cual es mandatorio de una biopsia renal percutánea con consecuentes eventos adversos.
Objetivo: Evaluar la capacidad que tienen los niveles de ADN libre circulante mitocondrial y nuclear urinario (uADN-lc) para discriminar muestras de sujetos con nefritis lúpica activa (NLA).
Sujetos y Métodos: Se tomaron muestras de orina de (74) sujetos, (25) con LES sin compromiso renal, (12) con NLA (12) con NLR y (25) Controles sanos. Se realizó una reacción en cadena de la polimerasa en tiempo real (qPCR) para la cuantificación de las moléculas de ácido desoxirribonucleico libre circulante (ADN-lc), mediante el empleo de genes constitutivos ND1 (mitocondrial) y GAPDH (nuclear), a posteriori de su extracción urinaria (uADN-lc), mediante el Kit Comercial de Aislamiento con Perlas Magnéticas (Applied Biosystems), y la determinación de su concentración con espectrofotómetro (Nanodrop One 2000 de Thermo Scientific ®) y analizador de imágenes.
Systemic lupus erythematous (SLE) is an autoimmune disease, prone to a heterogeneous clinical presentation that debuts silently in cases of active lupus nephritis (ALN), which is mandatory for a percutaneous renal biopsy with consequent adverse events. Objective: To evaluate the ability of mitochondrial and nuclear urinary cell-free DNA (cf-uDNA) levels to discriminate samples from subjects with active lupus nephritis (ALN). Subjects and Methods: Urine samples were taken from (74) subjects, (25) with SLE without kidney failure, (12) with (ALN), (12) with (RLN) and (25) Healthy Controls. A quantitative real-time polymerase chain reaction (Q-PCR) was performed for cell-free deoxyribonucleic acid molecule quantification (cf-DNA), using constitutive genes ND1 (mitochondrial) and GAPDH (nuclear), after their urinary extraction (cf-uDNA), by means of the Commercial Isolation Kit with Magnetic Pearls (Applied Biosystems), and its concentration measurement with spectrophotometer (Nanodrop One 2000 de Thermo Scientific ®) and image analyzer.
Systemic lupus erythematous (SLE) is an autoimmune disease, prone to a heterogeneous clinical presentation that debuts silently in cases of active lupus nephritis (ALN), which is mandatory for a percutaneous renal biopsy with consequent adverse events. Objective: To evaluate the ability of mitochondrial and nuclear urinary cell-free DNA (cf-uDNA) levels to discriminate samples from subjects with active lupus nephritis (ALN). Subjects and Methods: Urine samples were taken from (74) subjects, (25) with SLE without kidney failure, (12) with (ALN), (12) with (RLN) and (25) Healthy Controls. A quantitative real-time polymerase chain reaction (Q-PCR) was performed for cell-free deoxyribonucleic acid molecule quantification (cf-DNA), using constitutive genes ND1 (mitochondrial) and GAPDH (nuclear), after their urinary extraction (cf-uDNA), by means of the Commercial Isolation Kit with Magnetic Pearls (Applied Biosystems), and its concentration measurement with spectrophotometer (Nanodrop One 2000 de Thermo Scientific ®) and image analyzer.
Descripción
Palabras clave
Complejos inmunes, Concentración de ácido desoxirribonucleico libre circulante urinario (uADN-lc), GAPDH, SLEDAI, SLICC, ND1, Nefritis lúpica activa (NLA), Concentration of urinary circulating free deoxyribonucleic acid (uDNA-lc), Active lupus nephritis (ANL)