葡萄糖-6-磷酸脱氢酶缺乏症基因诊断新技术研究

目的：研究分析用于葡萄糖-6-磷酸脱氢酶（ glucose-6phosphate dehydrogenase ，G6PD）缺乏症基因诊断的多重SNaPshot基因诊断技术这一新技术的检出效果，以便于有效地进行新生儿筛查，和疾病的预防。方法选取2014年7月至2015年5月既往有G6PD生育史，或孕前筛查、新生儿筛查发现的该病患者和G6PD缺乏症高风险胎儿156例为研究对象，并将研究对象随机分为观察组和对照组，每组78例。对照组采用传统的基因检测技术G6PD／6-磷酸葡萄糖酸脱氢酶（6-phosphogluconate dehydrogenase ，6PGD）比值法，观察组则采用新的基因诊断技术-多重SNaP-shot基因诊断，分别检测对G6PD和编码该酶的基因诊断，以基因测序结果为“金标准”对比2组的检出结果。结果观察组待检者检出57例患有由基因突变导致的G6 PD缺乏症，总异常率为73?．08%（57／78），包括错义突变的A95 G型、G392T型、G1376T型、G1388A 型分别为5例、3例、17例、27例，其发生率分别为6．41%、3．85%、21．79%、34．62%，同义突变5例，发生率为6．41%，男、女异常率不同，女性均为杂合子；G6PD／6PGD比值法测定的78例待检者血样中总共有52例比值结果<1．0，总阳性率为66．67%（52／78），其中男性、女性的阳性例数分别为30、22；观察组采用的SNaPshot基因诊断技术，测出的突变类型及每种类型的突变数均与测序结果完全一致，而对照组采用的G6PD／6PGD比值法测定酶活性，只能测定错义突变，且存在漏诊，准确率为96．3%，无法检出同义突变。结论SNaPshot基因诊断技术可以有效地检出是那种基因突变导致G6PD缺乏，准确率高，有较高的临床推广应用价值。

An new diagnostic technique for gene diagnosis of glucose 6 phosphate dehydrogenase deficiency diseases

Objective To investigate the detection effect of multiple SNaPshot gene diagnosis technique in the diagbosis of glucose 6 phosphate dehydrogenase (G6PD) deficiency diseases in order to screen and prevent effectively neonatal diseases.Methods One hundred and fifty-six fetuses with G6PD deficiency diseases or with high risk of G6PD deficiency diseases who were diagnosed by progestation screening or neonatal screening in our hospital from July 2014 to May 2014 were enrolled in the study .The research objects were randomly divided into observation group and and control group ,with 78 fetuses in each group,the gene diagnosis for the coding gene of 6PGD was performed by traditional gene detection technique (specific value method) in control group or by multiple SNaPshot genetic diagnosis technique in observation group , respectively.The detection results were compared between two groups by taking gene sequencing results as “gold standard”.Results In observation group,57 patients with G6PD deficiency diseases that were caused by genetic mutation were detected ,with total abnormal rate being 73.085 (57/78), including missense mutation A95G, G392T, G1376T, G1388A,respectively in 5 cases, 3 cases, 17 cases,27 cases,respectively, with the incidence rate being 6.41%, 3.85%, 21.79%, 34.62%, respectively.Moreover there were 5 cases of samesense mutation , the incidence rate was 6.41%, and the abnormal rate was different between males and females that were all heterozygous .Among 78 blood samples detected by G6PD/6PGD specific value method,the specific value<1.0 in 52 cases,with total positive rate being 66.67%(52/78),in which the positive cases in male and female were 30 cases,22 cases,respectively.The mutation types and mutation number in every type detected by multiple SNaPshot genetic diagnosis technique were completely concordant with those detected by gene sequencing ,however, the enzymic activity detection by G6PD/6PGD specific value method could only determine missense mutation ,with accuracy being 96 .3%, however ,which could not detect samesense mutation .Conclusion The SNaPshot gene diagnosis technique can effectively detect the gene mutations which lead to G 6PD deficiency diseases ,with higher accuracy ,thus,which is worth using widely in clinical practice .