Screening And Detection Methods Of G6PD Deficiency In Newborns In S Of Zhongshan Area In The Past Ten Years And Analysis Of Genetic Testing Result Some Children

The study is divided into three pars.Part Ⅰ:Analysis of screening results of neonatal glucose-6-phosphate dehydrogenase deficiency in Zhongshan areaObjective:To investigate the relationship between gender,birth weight,gestational age and glucose-6-phosphate dehydrogenase deficiency(G6PDd)and G6PD population distribution in Zhongshan area,so as to guide clinical screening.Methods:Retrospective analysis of G6PDd screening results of 484014 neonates in Zhongshan area from 2010 to 2020 was performed by fluorescence analysis.Results:The positive rate of neonatal G6PDd screening was 4.474%(21653/484014),among which,the positive rate of male was 6.210%(16044/258405)and that of female was 2.482%(5597/225493).The positive rate of G6PDd screening in preterm infants was 4.753%(1122/23608),which was significantly higher than that in term infants 4.470%(19948/446257)and expired infants 3.348%(135/4032).The positive rate of G6PDd screening in giant infants was 3.502%(585/16705),significantly lower than 4.412%(990/22439)and 4.495%(990/22439)in low-birth-weight infants.Conclusion:Neonatal disease screening can effectively help the early diagnosis of G6PDd children,and preterm birth is an important factor affecting the activity of G6PD.In practice,clinicians can also take the distribution characteristics of NEONATAL G6PD activity into reference to do a more comprehensive screening of G6PDd.Part Ⅱ Evaluation of the performance of genetic screening processor and setting the cut-off values in screening for glucose-6-phosphate dehydrogenase disease in newbornsPurpose:The performance of Genetic Screening Processor(GSP)for determination of glucose-6-phosphate dehydrogenase(G6PD)in dried blood spots was verified,analyzed and evaluated.The cut-off value of glucose-6-phosphate dehydrogenase deficiency(G6PDd)screening by this method was established.Methods:The validation scheme was determined in the early stage,and the precision,accuracy and linear range of GSP system were verified for samples with B,D and E concentrations,quality control products with L and H concentrations and linear samples with 5 concentrations of kits.GSP analyzer and Victor2D 1420 fluorescence immunoassay(semi-automatic analyzer)were used to detect G6PD level in blood of 2733 neonates.Furthermore,percentile method and receiver Operating characteristic curve(ROC curve)method were used to establish the optimal cut-off value of G6PD level in blood of G6PDd screened by GSP analyzer in our laboratory.Results:When GSP was used to detect G6PD,the coefficient of variation in batch was 2.09%,1.54%and 1.90%,and the total coefficient of variation was 3.08%,1.72%and 2.12%,respectively,which were lower than 3.4%and 3.9%claimed by manufacturers.The relative deviation between the total mean of measurement and the target value of 25 samples of quality control concentration L was 3.84%,and that of 25 samples of quality control concentration H was 3.44%.The linear regression equation of tested values and theoretical values of five linear specimens was Y=1.0082x+0.0125,the square of correlation coefficient(R2)was 0.9991,R2>0.995,indicating good linear correlation.The G6PD level at the 5.0 percentile was 20.10 U/dL.Conclusion:GSP can be used in the detection of G6PD in zhongshan neonatal screening,and its precision,accuracy and linear range have reached the detection performance declared by the manufacturer,and it is routinely used in the screening of NEONATAL G6PD deficiency.The cut-off of GSP neonatal G6PD deficiency screening initially adopted by our center was 20.10 U/dL.Part Ⅲ:Genetic detection of newborn children with glucose-6-phosphate dehydrogenase deficiencyObjective:To investigate the gene mutation of newborn children with glucose-6-phosphate dehydrogenase deficiency in Zhongshan area,in order to better help clinical diagnosis.Methods:Neonates born in neonatal Disease Screening Center of Boai Hospital of Zhongshan city from June to July 2018 were selected as the research subjects.G6PD screening of neonatal dry blood plaque was performed by fluorescence analysis method.Blood card samples of 20 clinically confirmed CHILDREN with G6PDd were collected from the initial positive patients,and whole-genome DNA was extracted.Exon gene sequencing technology was used for sequencing analysis.Results:There were 21 clinically confirmed cases of G6PDd,including 16 males and 5 females.7 cases were c.1376G>T homozygous mutation,4 cases were c.1388G>A homozygous mutation,1 case was c.871G>A combined with c.13 76G>T heterozygous mutation,1 case was C.868-A homozygous combination c.1376G>T homozygous heterozygous mutation,1 case of c.1240-A combined with c.1376G>T heterozygous mutation,1 case of c.1311 C>T,c.1365-13T>C heterozygous mutation,1 case Case c.95A>G homozygous mutation,1 case c.95A>G hemizygous mutation,1 case c.871G>A homozygous mutation,1 case c.871G>A homozygous mutation,1 case c.835A>T Hemizygous mutation,1 case of c.392G>T homozygous mutation.Among them,the mutation frequency of c.1376G>T was the highest.Conclusion:C.1376G>T,C.1388G>A and C.95A>G are the three most common mutations of G6PD gene in Chinese population,which are all present in Chinese population,but have not been reported in other ethnic groups in the world.The results of this study were consistent with the common G6PD mutation sites in China,which enriched the G6PD gene map in Zhongshan area,provided genetic evidence for clinical diagnosis of glucose-6-phosphate dehydrogenase,and was more conducive to clinical work.