Rapid detection of solute carrier family 4, member 1 (SLC4A1) mutations and polymorphisms by high-resolution melting analysis

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• 作者：Choochai Nettuwakul ; Nunghathai Sawasdee ; Pa-thai Yenchitsomanus
• 关键词：Solute carrier family 4 member A ; SLC4A1 ; Anion exchanger 1 ; AE1 ; Band 3 ; Spherocytosis ; Ovalocytosis ; Distal renal tubular acidosis ; dRTA ; Real-time PCR ; Single nucleotide polymorphism ; SNP ; Mutation ; High-resolution melting
• 刊名：Clinical Biochemistry
• 出版年：2010
• 出版时间：March 2010
• 年：2010
• 卷：43
• 期：4-5
• 页码：497-504
• 全文大小：1872 K

文摘

Objective

The objective of this study is to develop and evaluate a high-resolution melting (HRM) method for detection of SLC4A1 mutations and polymorphisms.

Design and methods

The HRM method was optimized for detection of 18 known SLC4A1 variants. It was then used for analysis of 16 blind DNA samples highly enriched with two common mutations, Southeast Asian ovalocytosis (SAO) and band 3 Bangkok 1 (G701D), to compare the results with that of the conventional procedures.

Results

The HRM method was able to detect all 18 SLC4A1 variants. In the samples in which homozygous wild-type and homozygous variant could not be distinguished by difference plots, they were spiked with a sample carrying known homozygous genotype, resulting in their clear differentiation. The HRM method had 100 % efficiency for detection of mutations in the blind DNA samples, when compared with that of the conventional techniques.

Conclusions

The developed HRM method is efficient and reproducible for detection of SLC4A1 mutations and polymorphisms.