Mutation detection of ADPKD PKD1 gene in Hans by denaturing high-performance liquid chromatography.
- Author:
Shu-zhong ZHANG
1
;
Yu-hong ZHANG
;
Dian-yong ZHANG
;
Chang-lin MEI
Author Information
- Publication Type:Journal Article
- MeSH: Adult; Asian Continental Ancestry Group; genetics; Base Sequence; China; Chromatography, High Pressure Liquid; methods; DNA Mutational Analysis; Family Health; Female; Humans; Male; Middle Aged; Mutation; Pedigree; Polycystic Kidney, Autosomal Dominant; diagnosis; ethnology; genetics; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; TRPP Cation Channels; genetics
- From: Chinese Journal of Medical Genetics 2006;23(3):283-288
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop a screening system for more rapid and sensitive mutation detection of autosomal dominant polycystic kidney disease (ADPKD) gene 1 (PKD1) by using denaturing high-performance liquid chromatography (DHPLC) protocol.
METHODSUsing genomic DNA as templates extracted from blood samples of 19 Han pedigrees with 67 family members, the complete codon areas were amplified by long-range PCR and nested PCR in succession, and then the PCR products were analyzed by DHPLC. The mutations from screened abnormal PCR products were confirmed by DNA sequencing, and then compared with the mutations identified by single strand conformation polymorphism (SSCP) before.
RESULTSThere were 14 mutations found in this study, including 10 missense, 1 insertion, 1 deletion and 2 nonsense mutations. Besides 12 mutations identified before, mutations nt32819G>A and nt37137T>C were the novel mutations found. The mutation detection ratio was 73.7%.
CONCLUSIONThis developed system via DHPLC can be used as a more effective approach for mutation detection of autosomal dominant polycystic kidney disease PKD1 in Hans.