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1.
PLoS One ; 16(9): e0257350, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34555073

RESUMEN

SARS-CoV-2 has spread worldwide and has become a global health problem. As a result, the demand for inputs for diagnostic tests rose dramatically, as did the cost. Countries with inadequate infrastructure experience difficulties in expanding their qPCR testing capacity. Therefore, the development of sensitive and specific alternative methods is essential. This study aimed to develop, standardize, optimize, and validate conventional RT-PCR targeting the N gene of SARS-CoV-2 in naso-oropharyngeal swab samples compared to qPCR. Using bioinformatics tools, specific primers were determined, with a product expected to be 519 bp. The reaction conditions were optimized using a commercial positive control, and the detection limit was determined to be 100 fragments. To validate conventional RT-PCR, we determined a representative sampling of 346 samples from patients with suspected infection whose diagnosis was made in parallel with qPCR. A sensitivity of 92.1% and specificity of 100% were verified, with an accuracy of 95.66% and correlation coefficient of 0.913. Under current Brazilian conditions, this method generates approximately 60% savings compared to qPCR costs. Conventional RT-PCR, validated herein, showed sufficient results for the detection of SARS-CoV-2 and can be used as an alternative for epidemiological studies and interspecies correlations.


Asunto(s)
Prueba de Ácido Nucleico para COVID-19/métodos , COVID-19/diagnóstico , Nariz/virología , Proteínas de la Nucleocápside/genética , Orofaringe/virología , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , SARS-CoV-2/genética , Adolescente , Brasil , COVID-19/virología , Cartilla de ADN/genética , Femenino , Humanos , Masculino , Técnicas de Diagnóstico Molecular/métodos , ARN Viral/genética , Estándares de Referencia , Sensibilidad y Especificidad , Manejo de Especímenes/métodos
2.
Anim Reprod ; 15(2): 95-101, 2018 Aug 16.
Artículo en Inglés | MEDLINE | ID: mdl-34122638

RESUMEN

The study aimed at evaluating the testicular structure and spermatogenesis of Santa Inês and Santa Inês and Dorper crossbred sheep. Sixteen testicles of the animals under study were used, in order to evaluate the volumetric proportion, the diameter of the tubules, the height of seminiferous epithelium, the frequency of stages of the seminiferous epithelium cycle and the spermatogenic yield. The data were submitted to Student-Newman-Keulstestat 5% significance. The tubular diameter was 173.12 ± 29.09 µm and 185.71 ± 29.7 µm, and the height of the seminiferous epithelium was 52.29 ± 9.98 µm and 56.68 ± 11.25 µm, for the crossbred and the Santa Inês, respectively. There was no difference between the testicular compartments and the frequency of the stages of the seminiferous epithelium cycle between the groups. Santa Inês sheep presented a number of 15.36 ± 4.49 spermatocytes in the pre-leptotene/leptotene stage and 27.42 ± 6.65 spermatocytes in the pachytene, whereas the crossbred presented 13.18 ± 5.19 and 23.48 ± 7.80, respectively. The crossbred showed higher meiotic yield (3.98 ± 1.28) and spermatogenesis yield (3.71 ± 1.02). It is concluded there are differences in testicular morphology and spermatogenesis between Santa Inês sheep and crossbred Dorper/Santa Inês, indicating that the crossbreeding between the Santa Inês and Dorper animals allows a gain in the reproductive potential.

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