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.
Artículo
en Inglés
| LILACS
| ID: biblio-1415205
Biblioteca responsable:
BR18.1
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 sampleAsunto(s)
Texto completo:
Disponible
Colección:
Bases de datos internacionales
Base de datos:
LILACS
Asunto principal:
Reacción en Cadena de la Polimerasa
/
Tamizaje Neonatal
/
Análisis de Secuencia de ADN
Tipo de estudio:
Estudio diagnóstico
/
Guía de práctica clínica
/
Estudio de tamizaje
Límite:
Humanos
/
Recién nacido
Idioma:
Inglés
Revista:
Clin. biomed. res
Asunto de la revista:
Medicina
Año:
2022
Tipo del documento:
Artículo
País de afiliación:
Brasil
Institución/País de afiliación:
Hospital de Clínicas de Porto Alegre/BR
/
Universidade Federal do Rio Grande do Sul/BR