[Rapid diagnosis of non-deletion alpha-thalassemias by reverse dot blot].
OBJECTIVE
To establish a rapid and convenient method of reverse dot blot (RDB) analysis for detecting the point mutations of non-deletion alpha-thalassemia in Chinese.


METHODS
Label 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.


RESULTS
The 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.


CONCLUSIONS
This 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.
Li
Li-yan
LY
Department of Medical Genetics, First Military Medical University, Guangzhou, Guangdong, 510515 PR China.
Mo
Qiu-hua
QH
Xu
Xiang-min
XM
D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D017354:Point Mutation; D015203:Reproducibility of Results; D055542:alpha-Globins / Q000235:genetics; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics*
D055542:alpha-Globins