RESUMEN
Phylogenetic analysis has demonstrated that the etiologic agent of the 2020 pandemic outbreak is a betacoronavirus named SARS-CoV-2. For public health interventions, a diagnostic test with high sensitivity and specificity is required. The gold standard protocol for diagnosis by the Word Health Organization (WHO) is RT-PCR. To detect low viral loads and perform large-scale screening, a low-cost diagnostic test is necessary. Here, we developed a cost-effective test capable of detecting SARS-CoV-2. We validated an auxiliary protocol for molecular diagnosis with the SYBR Green RT-PCR methodology to successfully screen negative cases of SARS-CoV-2. Our results revealed a set of primers with high specificity and no homology with other viruses from the Coronovideae family or human respiratory tract pathogenic viruses, presenting with complementarity only for rhinoviruses/enteroviruses and Legionella spp. Optimization of the annealing temperature and polymerization time led to a high specificity in the PCR products. We have developed a more affordable and swift methodology for negative SARS-CoV-2 screening. This methodology can be applied on a large scale to soften panic and economic burden through guidance for isolation strategies.
Asunto(s)
Prueba de Ácido Nucleico para COVID-19/métodos , COVID-19/diagnóstico , Compuestos Orgánicos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , SARS-CoV-2 , Benzotiazoles , ADN de Cadena Simple , Diaminas , Enterovirus , Genoma Viral , Humanos , Filogenia , Reacción en Cadena de la Polimerasa , Quinolinas , Rhinovirus , Sensibilidad y Especificidad , Temperatura , Carga ViralRESUMEN
A expressão de RNAm para leptina, receptor de leptina (obRb), adiponectina, receptor de adiponectina (AdipoR1) e resistina foi avaliada por meio da técnica de PCR em tempo real, em tecidos ovariano, hipofisário, adiposo do omento e da região perirrenal, em ovelhas alimentadas sem farelo de mamona ou com farelo de mamona detoxificada durante 14 meses. O tipo de dieta não afetou os níveis de RNAm para leptina, obRb, adiponectina, AdipoR1 e resistina nos diferentes tecidos avaliados (P>0,05). Nos tecidos ovariano e hipofisário, não foi verificada a expressão da adiponecina e da resistina, respectivamente. Como consequência, pode-se concluir que o farelo de mamona detoxificada pode ser utilizado como fonte proteica na dieta de ovelhas, sem afetar a expressão do gene resistina e dos genes leptina e adiponectina, bem como de seus receptores...
The expression of leptin, leptin receptor (obRb), adiponectin, adiponectin receptor (AdipoR1) and resistin was assessed by real-time PCR technique in ovarian, pituitary, and the omental adipose perirenal tissue in sheep feed without castor meal or with detoxified castor meal. The type of diet did not affect mRNA levels for leptin, obRb, adiponectin, resistin AdipoR1 evaluated in different tissues (P>0.05). However, in pituitary and ovarian tissues there was no expression of resistin and adiponectin, respectively. The detoxified castor meal can be used in sheep diets as alternative food protein without affecting the expression of leptin and adponectin as well as their receptors and resistin...