Rapid detection of beta-thalassemia by LDR-ULP combined with real-time PCR / 中华检验医学杂志
Chinese Journal of Laboratory Medicine
; (12): 766-770, 2016.
Article
em Zh
| WPRIM
| ID: wpr-501808
Biblioteca responsável:
WPRO
ABSTRACT
Objective To establish a new method for rapid detection of β-thalassemia by investigating six clinical common mutation types.Methods Fifty cases of clinical wild-type samples and 42 cases ofβ-thalassemia samples were collected, and β-globin gene was amplified by PCR.Uniform ligation probe ( ULP) specific probes were designed for hybridization reaction to increase the reaction specificity and real-time PCR was performed to increase the sensitivity.After that, PCR products were verified by agarose electrophoresis.After examining the specificity and sensitivity, Kappa test between LDR-ULP method and reverse dot blot( RDB) method was conducted.Results Hybridization efficiency was improved 2.53 times by LDR-ULP hybridization.Each mutant type showed a significant amplification curve, whereas the wild-type had no significant curve within 40 cycles.The limit of determination of this method was 5 pg.The results of 92 cases of peripheral blood samples detected by the method of LDR-ULP and RDB were completely consistent.Conclusion In this study, a simple, inexpensive, rapid new method to detect β-thalassemia were established.
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Índice:
WPRIM
Tipo de estudo:
Diagnostic_studies
Idioma:
Zh
Revista:
Chinese Journal of Laboratory Medicine
Ano de publicação:
2016
Tipo de documento:
Article