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J Asthma ; 57(8): 850-857, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-31082286

RESUMEN

Objective: To develop a detection method for single nucleotide polymorphisms (SNPs) of bronchial asthma (BA) susceptibility genes (IL-13, IL-33, and GSDMA) based on fluorescence PCR melting curves.Methods: Peripheral blood samples from 33 patients with BA were collected. DNA was extracted, and positive plasmids were constructed. Probes and primers for fluorescence polymerase chain reaction (PCR) were designed according to IL-13, IL-33, and GSDMA sequences, and the SNPs were separately detected by gene sequencing and fluorescence PCR melting curve.Results: The system was successfully divided into 3 SNPs, including IL-13, IL-33, and GSDMA, and a comparison of sequencing methods showed that the results were completely consistent. The lowest detection limit was 1 ng/reaction, the sensitivity and specificity were 100%, and this method had high repeatability (CV = 2.8%).Conclusion: The fluorescence PCR melting curve method is suitable for the rapid and accurate classification of SNPs. The method is economical, simple, and efficient, and is suitable for the screening of the susceptible gene SNPs in a large-scale population of patients with BA.


Asunto(s)
Asma/diagnóstico , Pruebas Genéticas/métodos , Técnicas de Genotipaje/métodos , Tamizaje Masivo/métodos , Reacción en Cadena de la Polimerasa/métodos , Adolescente , Adulto , Asma/sangre , Asma/genética , Estudios de Factibilidad , Femenino , Fluorescencia , Predisposición Genética a la Enfermedad , Técnicas de Genotipaje/economía , Humanos , Interleucina-13/genética , Interleucina-33/genética , Masculino , Tamizaje Masivo/economía , Proteínas de Neoplasias/genética , Reacción en Cadena de la Polimerasa/economía , Polimorfismo de Nucleótido Simple , Sensibilidad y Especificidad , Adulto Joven
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