title: Rapid detection of G6PD mutations by multicolor melting curve analysis.
abstract: The MeltPro G6PD assay is the first commercial genetic test for glucose-6-phosphate dehydrogenase (G6PD) deficiency. This multicolor melting curve analysis-based real-time PCR assay is designed to genotype 16 G6PD mutations prevalent in the Chinese population. We comprehensively evaluated both the analytical and clinical performances of this assay. All 16 mutations were accurately genotyped, and the standard deviation of the measured Tm was <0.3°C. The limit of detection was 1.0ng/μL human genomic DNA. The assay could be run on four mainstream models of real-time PCR machines. The shortest running time (150min) was obtained with LightCycler 480 II. A clinical study using 763 samples collected from three hospitals indicated that, of 433 samples with reduced G6PD activity, the MeltPro assay identified 423 samples as mutant, yielding a clinical sensitivity of 97.7% (423/433). Of the 117 male samples with normal G6PD activity, the MeltPro assay confirmed that 116 samples were wild type, yielding a clinical specificity of 99.1% (116/117). Moreover, the MeltPro assay demonstrated 100% concordance with DNA sequencing for all targeted mutations. We concluded that the MeltPro G6PD assay is useful as a diagnostic or screening tool for G6PD deficiency in clinical settings.
authors: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China; United Diagnostic and Research Center for Clinical Genetics, School of Public Health of Xiamen University & Xiamen Maternal and Child Health Hospital, Xiamen, Fujian 361102, China.
authors: Zhongmin
authors: Z
authors: Xia
authors: Hemoglobin Laboratory, the First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, Guangxi Zhuang Autonomous Region, Nanning 530021,China.
authors: Ping
authors: P
authors: Chen
authors: Department of Clinical Laboratory, Liuzhou Key Laboratory of Birth Defects Prevention and Control, Liuzhou Maternal and Child Health Hospital, Liuzhou, Guangxi 545001, China.
authors: Ning
authors: N
authors: Tang
authors: Department of Clinical Laboratory, Liuzhou Key Laboratory of Birth Defects Prevention and Control, Liuzhou Maternal and Child Health Hospital, Liuzhou, Guangxi 545001, China.
authors: Tizhen
authors: T
authors: Yan
authors: Zhuhai Institute of Medical Genetics & Department of Clinical Laboratory, Zhuhai Municipal Maternity and Child Healthcare Hospital, Zhuhai, Guangdong 519001, China.
authors: Yuqiu
authors: Y
authors: Zhou
authors: Zhuhai Institute of Medical Genetics & Department of Clinical Laboratory, Zhuhai Municipal Maternity and Child Healthcare Hospital, Zhuhai, Guangdong 519001, China.
authors: Qizhi
authors: Q
authors: Xiao
authors: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China. Electronic address: hqying@xmu.edu.cn.
authors: Qiuying
authors: Q
authors: Huang
authors: State Key Laboratory of Cellular Stress Biology, State Key Laboratory of Molecular Vaccinology and Molecular Diagnostics, Engineering Research Centre of Molecular Diagnostics, Ministry of Education, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China. Electronic address: qgli@xmu.edu.cn.
authors: Qingge
authors: Q
authors: Li
mesh_terms: D005260:Female; D005820:Genetic Testing; D005838:Genotype; D005954:Glucosephosphate Dehydrogenase / Q000097:blood* / Q000235:genetics; D005955:Glucosephosphate Dehydrogenase Deficiency / Q000175:diagnosis* / Q000235:genetics / Q000473:pathology; D006801:Humans; D008297:Male; D009154:Mutation; D009691:Nucleic Acid Denaturation / Q000235:genetics*; D017422:Sequence Analysis, DNA
chemical_list: D005954:Glucosephosphate Dehydrogenase
keywords: Glucose-6-phosphate dehydrogenase deficiency; Multicolor melting curve analysis; Newborn screening; Real-time PCR