Development of the Next Generation Sequencing-Based Diagnostic Test for ß-Thalassemia and its Validation in a Pashtun Family.
Hemoglobin
; 44(4): 254-258, 2020 Jul.
Article
em En
| MEDLINE
| ID: mdl-32691635
ABSTRACT
ß-Thalassemia (ß-thal) is a common monogenic disease with ethnic-specific mutations on the HBB gene throughout the world. The reported mutations either reduce the expression or completely inactivate the HBB gene. In Pakistan, the prevalence of ß-thal is high due to consanguineous marriages. Accurate identification of mutations in carriers is imperative for prevention of ß-thal in subsequent generations. To overcome the limitations of traditional testing methods for ß-thal, a next-generation sequencing (NGS)-based diagnostic test was designed and validated by sequencing the entire HBB gene. The primer set covering the entire HBB gene was designed and validated in a Pashtun ß-thalassemic family. The polymerase chain reaction (PCR) product was sequenced using an Illumina MiSeq platform. A homozygous pathogenic insertion of A>AC/AC (rs35699606) was detected in an affected member of the family, while unaffected members were heterozygous for it. In addition, all family members were homozygous for the synonymous variant, A>G/G (rs713040), except the father who was heterozygous for it. We sequenced the entire HBB gene using the NGS-based test, which is highly sensitive, robust and specific for the diagnosis and screening of ß-thal in Pakistan, especially for families practicing consanguineous marriages.
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Texto completo:
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Base de dados:
MEDLINE
Assunto principal:
Testes Genéticos
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Talassemia beta
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Globinas beta
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Sequenciamento de Nucleotídeos em Larga Escala
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Mutação
Tipo de estudo:
Diagnostic_studies
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Prognostic_studies
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Risk_factors_studies
Limite:
Humans
País como assunto:
Asia
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article