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
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Fecha
2022
Autores
Márquez Escobar, Olga Cecilia
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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.
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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)
