Open AccessRapid screening of MMACHC gene mutations by high‐resolution melting curve analysis
Author(s) -
Wang Chao,
Liu Yang,
Cai Fengying,
Zhang Xinjie,
Xu Xiaowei,
Li Yani,
Zou Qianqian,
Zheng Jie,
Zhang Yuqin,
Guo Wei,
Cai Chunquan,
Shu Jianbo
Publication year - 2020
Publication title -
molecular genetics and genomic medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.765
H-Index - 29
ISSN - 2324-9269
DOI - 10.1002/mgg3.1221
Subject(s) - methylmalonic acidemia , high resolution melt , genetics , sanger sequencing , cobalamin , melting curve analysis , biology , population , mutation , medicine , gene , polymerase chain reaction , environmental health , vitamin b12 , endocrinology
Background Cobalamin (cbl) C is a treatable rare hereditary disorder of cbl metabolism with autosomal recessive inheritance. It is the most common organic acidemia, manifested as methylmalonic academia combined with homocysteinemia. Early screening and diagnosis are important. The mutation spectrum of the MMACHC gene causing cblC varies among populations. The mutation spectrum in Chinese population is notably different from that in other populations. Methods A PCR followed by high‐resolution melting curve analysis (PCR‐HRM) method covering all coding exons of MMACHC gene was designed to verify 14 pathogenic MMACHC gene variants found in patients with cblC, including all common mutations in Chinese patients with cblC. Result By PCR‐HRM analysis, 14 pathogenic variants of MMACHC showed distinctly different melting curves, which were consistent with Sanger sequencing. The homozygous type of the most common mutation c.609G > A (p.Trp203Ter) can also be analyzed by specially designed PCR‐HRM. Conclusion The established PCR‐HRM method for screening common pathogenic MMACHC variants in Chinese patients with cblC has the advantages of high accuracy, high throughput, low cost, and high speed. It is suitable for the large‐sample screening of suspected children with methylmalonic acidemia and carriers in population.