1. A novel high-resolution melting analysis strategy for detecting cystic fibrosis–causing variants.
- Author
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Rodríguez, Gerardo Raúl Díez, Lago, Juan Emilio Figueredo, Cayarga, Anny Armas, González, Yaimé Josefina González, Rosa, Iria García de la, Mesa, Teresa Collazo, Reyes, Ixchel López, Lozada, Yulaimy Batista, Calá, Fidel Ramón Rodríguez, and Sánchez, Juan Bautista García
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CYSTIC fibrosis diagnosis , *DNA analysis , *FREEZING , *POLYMERASE chain reaction , *GENES , *GENETIC polymorphisms , *GENETIC mutation , *CYSTIC fibrosis , *ALLELES , *MEMBRANE proteins - 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. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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