Rapid diagnosis of non-deletion alpha-thalassemias by reverse dot blot.
- Author:
Li-yan LI
1
;
Qiu-hua MO
;
Xiang-min XU
Author Information
- Publication Type:Journal Article
- MeSH: Humans; Nucleic Acid Hybridization; methods; Point Mutation; Reproducibility of Results; alpha-Globins; genetics; alpha-Thalassemia; diagnosis; genetics
- From: Chinese Journal of Medical Genetics 2003;20(4):345-347
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a rapid and convenient method of reverse dot blot (RDB) analysis for detecting the point mutations of non-deletion alpha-thalassemia in Chinese.
METHODSLabel biotin to primers and amplify human alpha2 globin gene selectively, then hybridize PCR products with a set of oligonucleotide probes immobilized on strips, and develop colour to detect non-deletion alpha-thalassemia defects.
RESULTSThe PCR system using biotin-labeled primers could specifically amplify a 1085 bp fragment of human alpha2 globin gene which encompasses all six alpha-thalassemia mutations. After being hybridized with sequence-specific oligonucleotide probes and colour development, it could simultaneously identify all six types of non-deletion alpha-thalassemias encountered in Chinese.
CONCLUSIONThis method does not need semi-nested PCR, and the products amplified by biotinylated primers can be used directly to hybridize with the probes on strips under the identical conditions of hybridization. So, it is a specific and multiplex detection assay for screening non-deletion alpha-thalassemia defects in Chinese.