Standardization of an organic DNA extraction method from dried blood spots and its downstream molecular applications in neonatal screening and diagnostic confirmation of lysosomal disorders
Clin. biomed. res
;
42(3): 218-225, 2022.
Artigo
em Inglês
| LILACS
| ID: biblio-1415205
ABSTRACT
Introduction:
Dried blood spot (DBS) samples have been used for diagnostic purposes since their introduction in the neonatal screening of phenylketonuria almost 50 years ago. The range of its application has been extended to modern approaches, such as next-generation sequencing (NGS) for molecular genetic testing. This study aimed to evaluate the use of a standardized organic method for DNA extraction from DBS samples in the diagnostic setting.Methods:
The clinical applicability of the method was tested using 3 samples collected from a newborn screening project for lysosomal storage diseases, allowing the determination of the genotype of the individuals. DNA was extracted from 3 3-mm diameter DBS punches. Quality, purity, and concentration were determined, and method performance was assessed by standard polymerase chain reaction, restriction length polymorphism, Sanger sequencing, and targeted NGS.Results:
Results were compared with the ones obtained from DNA samples extracted following the internally validated in-house extraction protocol that used 6 3-mm punches of DBS and samples extracted from whole blood.Conclusion:
This organic method proved to be effective in obtaining high-quality DNA from DBS, being compatible with several downstream molecular applications, in addition to having a lower cost per sampleAssuntos
Texto completo:
DisponíveL
Índice:
LILACS (Américas)
Assunto principal:
Reação em Cadeia da Polimerase
/
Triagem Neonatal
/
Análise de Sequência de DNA
Tipo de estudo:
Estudo diagnóstico
/
Guia de Prática Clínica
/
Estudo de rastreamento
Limite:
Humanos
/
Recém-Nascido
Idioma:
Inglês
Revista:
Clin. biomed. res
Assunto da revista:
Medicina
Ano de publicação:
2022
Tipo de documento:
Artigo
País de afiliação:
Brasil
Instituição/País de afiliação:
Hospital de Clínicas de Porto Alegre/BR
/
Universidade Federal do Rio Grande do Sul/BR
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