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Cystic Fibrosis: A Simple and Customized Strategy for Genetic Screening Able to Detect Over 90% of Identified Mutated Alleles in Brazilian Newborns

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Abstract

Introduction

The incorporation of molecular genetic testing into cystic fibrosis (CF) screening programs increases the specificity of the diagnostic strategy and has the potential to decrease the rate of false- positive results. In this sense, our objective was to develop a genotyping assay that could detect 25 pathogenic variants in the cystic fibrosis transmembrane conductance regulator (CFTR) gene with high sensitivity and that could be incorporated into the routine of newborn screening, complementing the current existing protocol used in our public health institution.

Methods

A mini-sequencing assay was standardized using single-base extension in a previously genotyped control sample. This strategy was validated in a Brazilian cohort of CF patients by Sanger sequencing.

Results

The inclusion of the 25 variants in the current newborn screening program increased the identification rates of two alleles from 33 to 52.43% in CF patients. This new approach was able to detect a total of 37 variants, which represents 93.01% of all mutated alleles described in the last CF Brazilian Register.

Conclusions

Mini-sequencing for the simultaneous detection of 25 CFTR gene variants improves the screening of Brazilian newborns and decreases the number of inconclusive cases. This method uses minimal hands-on time and is suited for rapid screening, which reduces sample processing costs.

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Author information

Author notes

  1. José Eduardo Vargas and Maria L. R. Rossetti contributed equally to this work.

Authors and Affiliations

  1. Programa de Pós-Graduação em Biologia Celular e Molecular, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Thaiane Rispoli, Mayara J. Prado & Maria L. R. Rossetti

  2. Centro de Desenvolvimento Científico e Tecnológico, Secretaria da Saúde do Estado do Rio Grande do Sul (SES-RS), Porto Alegre, Rio Grande do Sul, Brazil

    Thaiane Rispoli, Grazielle M. Rodrigues, Mayara J. Prado, Cláudia M. D. da Silva & Tarciana Grandi

  3. Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Grazielle M. Rodrigues & Simone M. de Castro

  4. Serviço de Referência em Triagem Neonatal, Hospital Materno Infantil Presidente Vargas, Porto Alegre, Rio Grande do Sul, Brazil

    Simone M. de Castro & Helena T. Mocelin

  5. Hospital da Criança Santo Antônio, Irmandade da Santa Casa de Misericórdia de Porto Alegre, Porto Alegre, Rio Grande do Sul, Brazil

    Gilberto B. Fischer

  6. Hospital São Lucas da PUCRS, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Leonardo A. Pinto

  7. Hospital de Clínicas de Porto Alegre, Porto Alegre, Rio Grande do Sul, Brazil

    Paulo J. C. Maróstica

  8. Programa de Pós-Graduação em Ciências Pneumológicas, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Laís C. R. Scortegagna

  9. Instituto de Ciências Biológicas, Universidade de Passo Fundo, Passo Fundo, Rio Grande do Sul, Brazil

    José Eduardo Vargas

  10. Programa de Pós-Graduação em Biologia Molecular Aplicada a Saúde, Canoas, Universidade Luterana do Brazil, Canoas, Rio Grande do Sul, Brazil

    Maria L. R. Rossetti

Authors
  1. Thaiane Rispoli
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  2. Grazielle M. Rodrigues
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  3. Simone M. de Castro
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  4. Mayara J. Prado
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  5. Cláudia M. D. da Silva
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  6. Tarciana Grandi
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  7. Gilberto B. Fischer
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  8. Leonardo A. Pinto
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  9. Paulo J. C. Maróstica
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  10. Laís C. R. Scortegagna
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  11. Helena T. Mocelin
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  12. José Eduardo Vargas
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  13. Maria L. R. Rossetti
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Corresponding authors

Correspondence to Thaiane Rispoli or José Eduardo Vargas.

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Funding

The study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) (Finance Code 001) and Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) (1260-2551/13-0).

Ethics approval

This study was approved by the ethics committee of the Secretaria da Saúde do Estado do Rio Grande do Sul, Universidade Federal do Rio Grande do Sul, Hospital Materno Infantil Presidente Vargas, Hospital da Criança Santo Antônio, Hospital São Lucas da PUCRS, and Hospital de Clínicas de Porto Alegre.

Patient consent

Obtained.

Conflict of interest

The authors, Thaiane Rispoli, Grazielle M. Rodrigues, Simone M. de Castro, Mayara J. Prado, Cláudia M.D. da Silva, Tarciana Grandi, Gilberto B Fischer, Leonardo A. Pinto, Paulo J.C. Maróstica, Laís C.R. Scortegagna, Helena T. Mocelin, José E. Vargas, and Maria L.R. Rossetti, have no conflict of interest to declare.

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Rispoli, T., Rodrigues, G.M., de Castro, S.M. et al. Cystic Fibrosis: A Simple and Customized Strategy for Genetic Screening Able to Detect Over 90% of Identified Mutated Alleles in Brazilian Newborns. Mol Diagn Ther 24, 315–325 (2020). https://doi.org/10.1007/s40291-020-00456-9

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  • DOI: https://doi.org/10.1007/s40291-020-00456-9

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