Original Article
The clinical value of fluorescence quantitative assay and G6PD/6PGD ratio method in the diagnosis of glucose-6-phosphate dehydrogenase deficiency and gene mutation
Zhang Xiaofen, Tian Guoli, Ji Wei, Zhou Zhuo, Guo Jing, Wang Yanmin
Published 2022-05-19
Cite as Chin J Lab Med, 2022, 45(5): 522-527. DOI: 10.3760/cma.j.cn114452-20210927-00606
Abstract
ObjectiveTo investigate the diagnostic value of fluorescence quantitative method and G6PD/6PGD ratio method in glucose-6-phosphate dehydrogenase (G6PD) deficiency and the type of gene mutation.
MethodsA total of 1 201 patients (711 males and 490 females) with suspected G6PD deficiency in Shanghai Children′s Hospital were collected from June 2018 to March 2021. Fluorescence quantification method, G6PD/6PGD ratio method and multicolor melting curve were used to detects enzyme activity, ratio and gene mutation type. Comparison of each index and evaluation of its diagnostic efficiency were performed.
ResultsAmong 1 201 suspicious samples, 163 cases (135 males and 28 females) were finally diagnosed. 156 cases were diagnosed by fluorescence quantitative method with a detection rate of 95.71%, and 140 cases were diagnosed by G6PD/6PGD ratio method with a detection rate of 85.89%. enzymatic activity of G6PD and ratio of G6PD/6PGD in male were significantly lower than female, and the differences were statistically significant (U=642.5, 734.5, P<0.001). 112 cases received G6PD gene mutation detection and 92 cases were diagnosed, 74 were hemizygous mutations, 1 were homozygous mutations, 15 were heterozygous mutations, and 2 were compound heterozygous mutations. Among 15 cases of heterozygous mutations, 11 cases were diagnosed by fluorescence quantitative method, the diagnosed rate was 73.33%, 4 cases were diagnosed by G6PD/6PGD ratio method, and the diagnosed rate was 26.67%. A total of 7 mutation sites were detected and the proportions were c.1388G>A (32.22%), c.1376G>T (30.00%), c.871G>A (13.33%), c.1024C>T (11.11%). c.95A>G (7.78%), c.487G>A (4.44%), c.392G>T (1.11%). The enzymatic activities of c.1376G>T and c.1024C>T, c.487G>A were statistically significant (P<0.001,0.015); the G6PD/6PGD ratios of c.1024C>T and c.1388G>A, c.1376G>T were statistically significant (P=0.017,0.002,0.011,0.013). Fluorescence quantitative method had sensitivity of 100%, specificity of 95.65%, and the area under the curve (AUC) is 0.972. The sensitivity of the G6PD/6PGD ratio method was 100%, the specificity was 94.57%, and the AUC was 0.979. The sensitivity of fluorescence quantitative method combined with G6PD/6PGD ratio was 96.7%, the specificity was 100%, and the AUC was 0.992.
ConclusionsCompared with fluorescence quantification, the G6PD/6PGD ratio method might not be able to diagnose female heterozygotes effectively; The panel of G6PD fluorescence quantification and G6PD/6PGD ratio was helpful to reduce the missed diagnosis. Combined with gene mutation analysis, it could improve the diagnosis rate of G6PD deficiency in the children.
Key words:
Glucose-6-phosphate dehydrogenase deficiency; Fluorescence quantitative assay; G6PD/6PGD ratio Method; Multicolor melting curve analysis; Gene mutation
Contributor Information
Zhang Xiaofen
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
Tian Guoli
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
Ji Wei
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
Zhou Zhuo
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
Guo Jing
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
Wang Yanmin
Department of Newborn Screening Center, Shanghai Children′ s Hospital, School of Medicine ShanghaiJiao tong University, Shanghai 200040, China
