High-throughput beta-thalassemia carrier screening by allele-specific Q-primer real-time polymerase chain reaction.
Anal Biochem
; 404(1): 97-9, 2010 Sep 01.
Article
en En
| MEDLINE
| ID: mdl-20433808
ABSTRACT
Based on a novel Q-primer real-time polymerase chain reaction (PCR) system, we designed allele-specific Q-primers for the detection of three beta-thalassemia mutations [Cd41/42(-TCTT), IVSI nt5 (G>C), and IVSII nt654 (C>T)] that have a high carrier frequency in Southeast Asia. With clear distinction between heterozygote and wild-type, DeltaC(t) (threshold cycle) values were defined. The results of evaluating 139 blinded samples by our system match perfectly with those obtained by the conventional reverse dot blot (RDB) method. With a 384-well plate that included replicates in the same analysis, our throughput reached 190 reactions per run with a turnaround time as short as 130 min, and the cost of consumables was as low as $1 (US) for each test.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Reacción en Cadena de la Polimerasa
/
Talasemia beta
/
Cartilla de ADN
/
Ensayos Analíticos de Alto Rendimiento
/
Tamización de Portadores Genéticos
Tipo de estudio:
Diagnostic_studies
/
Screening_studies
Límite:
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
2010
Tipo del documento:
Article
País de afiliación:
Singapur