RESUMO
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.
Assuntos
Asma/diagnóstico , Testes Genéticos/métodos , Técnicas de Genotipagem/métodos , Programas de Rastreamento/métodos , Reação em Cadeia da Polimerase/métodos , Adolescente , Adulto , Asma/sangue , Asma/genética , Estudos de Viabilidade , Feminino , Fluorescência , Predisposição Genética para Doença , Técnicas de Genotipagem/economia , Humanos , Interleucina-13/genética , Interleucina-33/genética , Masculino , Programas de Rastreamento/economia , Proteínas de Neoplasias/genética , Reação em Cadeia da Polimerase/economia , Polimorfismo de Nucleotídeo Único , Sensibilidade e Especificidade , Adulto JovemRESUMO
Primary hyperparathyroidism (PHPT) is characterized by inappropriate secretion of parathyroid hormone (PTH). The proportion of PHPT with asymptomatic or nontypical symptoms is continuously increasing. PHPT typically affects elderly women, and it is characterized by few symptoms or complications. We explored a case of a 77-year-old woman with previously undiagnosed PHPT who presented first with palpitation and chest distress, and then had the symptoms of intractable nausea and vomiting. The initial laboratory investigations revealed normal calcium levels. Further examination showed serum calcium and PTH were elevated. In order to attach importance to PHPT in asymptomatic patients or patients with nontypical symptoms, related literature was reviewed. Finally the methods of treatment of PHPT in elderly patients were discussed.
