A novel high-resolution melting analysis strategy for detecting cystic fibrosis-causing variants.

Laboratory medicine Pub Date : 2024-03-07 DOI:10.1093/labmed/lmad058

Gerardo Raúl Díez Rodríguez, Juan Emilio Figueredo Lago, Anny Armas Cayarga, Yaimé Josefina González González, Iria García de la Rosa, Teresa Collazo Mesa, Ixchel López Reyes, Yulaimy Batista Lozada, Fidel Ramón Rodríguez Calá, Juan Bautista García Sánchez

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Abstract

Cystic fibrosis (CF), an autosomal recessive disease, is caused by variants in both alleles of the CF transmembrane conductance regulator (CFTR) gene. A new assay based on allele-specific polymerase chain reaction and high-resolution melting analysis was developed for the detection of 18 CF-causing CFTR variants previously identified in Cuba and Latin America. The assay is also useful for zygosity determination of mutated alleles and includes internal controls. The reaction mixtures were normalized and evaluated using blood samples collected on filter paper. The evaluation of analytical parameters demonstrated the specificity and sensitivity of the method to detect the included CFTR variants. Internal and external validations yielded a 100% agreement between the new assay and the used reference tests. This assay can complement CF newborn screening not only in Cuba but also in Latin America.

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用于检测囊性纤维化致病变体的新型高分辨率熔融分析策略。

囊性纤维化（CF）是一种常染色体隐性遗传病，由 CF 跨膜传导调节器（CFTR）基因的两个等位基因变异引起。我们开发了一种基于等位基因特异性聚合酶链反应和高分辨率熔解分析的新测定法，用于检测之前在古巴和拉丁美洲发现的 18 种导致 CF 的 CFTR 变异基因。该检测方法还可用于突变等位基因的等位基因测定，并包括内部对照。使用滤纸收集的血样对反应混合物进行了归一化和评估。对分析参数的评估证明了该方法在检测 CFTR 变异方面的特异性和灵敏度。内部和外部验证结果表明，新检测方法与使用的参考检测方法之间的一致性达到了 100%。这种检测方法不仅可以补充古巴的 CF 新生儿筛查，还可以补充拉丁美洲的 CF 新生儿筛查。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Laboratory medicine

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