PAH基因点突变及IgG亚型异常与新生儿免疫代谢病的相关性研究

发布时间：2018-04-15 09:08

本文选题：PAH基因突变 + IgG亚型　；参考：《青岛大学》2017年硕士论文

【摘要】：目的通过对威海地区2013年至2015年出生的69110例新生儿进行筛查,以探讨威海地区新生儿免疫代谢病的发病情况及与其他省市地区的差异,重点观测PAH基因突变的类型及IgG抗体亚型异常与新生儿免疫代谢病相关性。对孕期的用药指导和确诊后的及时有效治疗,降低出生缺陷具有重要价值。方法自2013年～2015年期间,共有69110例新生儿在威海出生,并且经过家长同意接受新生儿疾病筛查的检查。通过采用时间分辨荧光技术和串联质谱技术对这些新生儿的血液样本进行筛查,回顾性的分析这三年间威海地区CH(甲状腺功能减低症)、PKU(苯丙酮尿症)、CAH(肾上腺皮质增生症)、G6PD(葡萄糖-6-磷酸脱氢酶缺乏症)及IMD(遗传代谢病)的发病率以及确诊患儿的治疗情况。同时采用PCR扩增技术对PKU患儿的PAH基因的13个外显子的扩增产物进行基因测序分析。采用ELISA对2013～2015年来威海市妇幼保健院就诊的母亲为O型血的354例发生新生儿溶血病的患儿及其母亲血清内的IgG亚型进行定量分析,同时利用100例健康的O型孕妇及100例健康的新生儿作为对照。结果1.威海地区2013-2015年间的年平均筛查率为99.71%,共计确诊CH患儿75例,发病率为1/921;PKU患儿11例,发病率为1/6283;CAH患儿5例,发病率为1/13822;G6PD患儿25例,发病率为1/2764;IMD患儿12例,发病率为1/2982。2.威海地区CH的发病率1/921高于全国的平均发病率1/3009,差异有统计学意义(P0.01)。3.通过对PKU患儿的基因检测发现14种54个基因突变,其中E7为突变高频区,患儿基因以杂合突变为主,PAH基因与新生儿遗传代谢性疾病PKU密切相关。通过基因手段对患儿及其父母进行检测,分析相关影响因素,对降低出生缺陷具有重要价值。4.IgG1和IgG3的含量在新生儿溶血病患儿母亲血清中明显高于正常母亲组(P0.05),IgG1和IgG3在患儿体内的含量也明显高于健康新生儿组(P0.05)。结论威海地区新筛筛查率很高,主要是有政府资金的大力支持,实行免费筛查。通过新生儿疾病筛查的检测手段能够做到早期发现、早期治疗。威海地区CH发病率高与孕期补碘过量关系密切。通过PCR扩增及基因检测手段对PKU患儿的突变基因进行明确的诊断,发现高频突变位点,对以后的PAH基因突变检测有很好的指导作用,同时可对有再生育需求的家庭进行孕期指导和产前诊断。用ELISA法检测母体及新生儿体内的IgG亚型的含量对新生儿溶血病的诊断以及判断预后有着重大意义,同时对孕期的临床用药也有很好的指导作用,有效的降低出生缺陷。
[Abstract]:Objective to screen 69110 newborns born in Weihai from 2013 to 2015, and to explore the incidence of neonatal immune metabolic disease and its difference from other provinces and cities in Weihai.The type of PAH gene mutation and the abnormality of IgG antibody subtype were observed.It is of great value to guide the use of drugs during pregnancy and timely and effective treatment after diagnosis to reduce birth defects.Methods from 2013 to 2015, a total of 69110 newborns were born in Weihai.Blood samples of these newborns were screened by time-resolved fluorescence and tandem mass spectrometry.The incidence of CH-PKU (phenylketonuria) and the incidence of G6PDand IMD (glucose-6-phosphate dehydrogenase deficiency) in Weihai area during the past three years were analyzed retrospectively.At the same time, 13 exons of PAH gene were sequenced by PCR.The serum IgG subtypes of 354 neonates with hemolytic disease of newborns and their mothers were quantitatively analyzed by ELISA from 2013 to 2015 in Weihai Maternal and Child Health Care Hospital.At the same time, 100 healthy type O pregnant women and 100 healthy newborns were used as control group.Result 1.The average annual screening rate in Weihai area from 2013 to 2015 was 99.71, with a total of 75 cases of confirmed Ch, 11 cases of 1 / 921 PKU, 5 cases of 1 / 6283 cases of CAH, 25 cases of 1 / 1 13822 cases of G6PD and 12 cases of 1 / 2764 IMD, with an incidence of 1 / 2982.2%.The incidence of Ch in Weihai area is 1 / 921 higher than the national average incidence rate of 1 / 3009, the difference is statistically significant.14 kinds of 54 gene mutations were found in children with PKU, of which E7 was the high frequency region of mutation. Heterozygosity was the main gene mutation in children with PKU and PKU was closely related to neonatal metabolic disease.The children and their parents were detected by genetic methods, and the related influencing factors were analyzed.4. The levels of IgG1 and IgG3 in mothers with neonatal hemolytic disease were significantly higher than those in normal mothers. The levels of IgG1 and IgG3 in children with neonatal hemolytic disease were also significantly higher than those in healthy newborns.Conclusion the screening rate of new screen in Weihai area is very high, which is mainly supported by government funds and free screening.Early detection and early treatment can be achieved by means of neonatal disease screening.The high incidence of Ch in Weihai area is closely related to iodine overdose during pregnancy.By means of PCR amplification and gene detection, the mutation genes of children with PKU were clearly diagnosed, and the high frequency mutation sites were found, which could guide the detection of PAH gene mutation in the future.Pregnancy guidance and prenatal diagnosis can also be given to families with reproductive needs.The detection of IgG subtype in mother and newborn by ELISA method is of great significance in the diagnosis and prognosis of hemolytic disease of newborn. It is also a good guide to the clinical use of drugs during pregnancy and can effectively reduce birth defects.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R722.1

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