母儿ABO血型不合的产前诊断与治疗

陕西中医学院硕士学位论文母儿ABO血型不合的产前诊断与治疗姓名：刘娜申请学位级别：硕士专业：中西医结合临床指导教师：景茹草20100401母儿ABO血型不合的产前诊断与治疗摘要母儿血型不合是指孕妇与胎儿间因血型不合产生的同族血型免疫性疾病，是新生儿溶血性疾病(hemolytic disease of the newborn,HDN)的重要病因，轻者症状不明显或出现黄疸，严重者中枢神经损伤，发生胎儿水肿、死胎、死产、新生儿胆红素脑病，导致脑瘫，甚至死亡。

Rh血型、ABO血型和MN血型不符均能引起这种疾病，在我国以ABO血型不合溶血病发生率最高，约占母儿血型不合的85.3%，Rh血型不合溶血病发生较少，MN溶血最为罕见。

传统用于产前诊断的材料一般通过羊水和绒毛膜穿刺等损伤性手段获取，这些方法易引起感染、流产。

1997年，Lo等[1]证实在孕妇外周血中存在相对较为丰富的游离胎儿DNA，这一发现为无创性产前诊断开辟了新的途径。

几年来，国内外学者利用母血循环中游离胎儿DNA，对各种胎儿疾病和病理性妊娠进行研究。

本课题提取O型血孕妇血浆中的游离胎儿DNA，采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法检测胎儿ABO血型，对母儿ABO血型不合患者进行产前诊断。

中医中药在治疗母儿血型不合方面，疗效满意, 较现代医学有着独特的优势。

我们所采用的中西医结合治疗方法可有效地预防胎儿及新生儿溶血所致并发症的发生，提高新生儿生存质量，是一种较理想的治疗产前母儿 ABO溶血的方法,故可在临床推广运用，对预防新生儿溶血病的发生有着重要的意义。

【目的】通过提取O型血孕妇血浆中的游离胎儿DNA，应用PCR-RFLP法提前检测胎儿ABO 血型的研究，来寻找一条快速、安全、灵敏度高、特异性强的非损伤性产前诊断母儿ABO血型不合的新方法。

对母儿ABO血型不合患者给予中西医结合治疗，以达到治疗母儿ABO血型不合，预防和降低新生儿溶血病的发病率和死亡率的目的。

【方法】使用硅胶膜的方法从丈夫血型为非O型的孕13～37周O型血健康单胎孕妇的血浆中提取出胎儿游离 DNA，然后采用PCR-RFLP法检测胎儿 ABO血型 ,分别扩增ABO 糖基转移酶基因中2个特异性片段，之后分别用限制性内切酶AluⅠ和KpnⅠ酶切消化，酶切产物电泳确定胎儿血型。

通过胎儿血型检测结果诊断母儿ABO血型不合。

对母儿ABO血型不合患者采用中西医结合的方法治疗。

中药处方为茵陈退黄汤，其中包括茵陈蒿12g、黄芩9g、大黄（制）4.5g、甘草6g；西药处方为维生素C注射液3g加入10%葡萄糖溶液500ml中，静脉滴注或50%葡萄糖溶液40ml加维生素C 注射液500mg静脉推注，每日1次，维生素E 100mg口服，每日1次，同时每日吸氧2次，每次30min，10天为一疗程。

【结果】在50例孕妇血浆中检测出胎儿血型32例，其中胎儿血型为A型者12例，B型者11例，O型者9例，检出率为64%，出生后血清学证实30例诊断正确，准确率为94%。

研究表明，孕妇外周血中确实存在着胎儿游离DNA，我们可以使用硅胶膜的方法将其提取出来；PCR-RFLP法检测胎儿 ABO血型是一种行之有效的无创性方法，值得推广应用；利用孕妇血浆中的游离胎儿 DNA可以产前诊断胎儿 ABO 血型，简便无损伤，对于诊断和预防新生儿溶血病的发生具有重要意义。

母儿ABO血型不合患者经过中西医结合方案治疗后，23例患者中，显效20例，有效3例，总有效率为100%。

这说明，中西医结合治疗方案是一种可以降低血清抗体效价，治疗母儿ABO血型不合，预防和减少新生儿溶血病的发病率和死亡率的有效方法。

【结论】利用孕妇血浆中游离胎儿DNA，能够检测胎儿ABO血型，产前诊断母儿ABO血型不合。

由此推测，母血循环中的游离胎儿DNA可能会对多种胎儿疾病和病理性妊娠的产前诊断有所帮助，它将有可能取代传统的产前诊断方法，成为一种新的无创性产前诊断方法，并逐渐应用于临床。

中西医结合治疗方案产前防治母儿ABO血型不合的疗效满意，此方法值得进行深入研究并推广。

关键词胎儿DNA；ABO血型；母儿ABO血型不合；中西医结合治疗Feto-Maternal ABO Blood Group Incompatibility of PrenatalDiagnosis and TreatmentAbstractThe Feto-Maternal Blood Group Incompatibility is the same family blood autoimmune diseases produced by blood group incompatibility between the pregnant women and fetus, is an important cause of hemolytic disease of the newborn (HDN). Light were less marked or jaundice. In severe cases, central nervous system has been damaged, cause Fetal Edema、Stillbirth or Newborn Bilirubin-encephalopathy,lead to cerebral palsy or even death. Rh blood group、ABO blood group and MN blood group can cause the disease.The hemolytic disease by ABO blood group incompatibility is the highest incidence in our country,about accounting for 85.3%.Rh blood group incompatibility hemolytic disease occurred in less. MN blood group incompatibility hemolytic disease is the most rare.Traditionally, materials used for prenatal diagnosis were got usually through invasive means such as amniotic and chorionic puncture. These methods easily cause infection and abortion.In 1997, Lo and so confirmed that there is relatively rich in free fetal DNA in the blood circulation of pregnant women . The discovery has opened up new avenues for non-invasive prenatal diagnosis.In recent years,domestic and foreign scholars have studied a variety of fetal diseases and pathological pregnancy in the use of free fetal DNA in maternal circulation. We extract the free fetal DNA in the O-type blood plasma of pregnant women，using polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) to detect fetal ABO blood group, for prenatal diagnosis the patients of Feto-Maternal ABO Blood Group Incompatibility .Medicine in the treatment of maternal-fetal blood group incompatibility,has content curative effect.It has a uniquer advantage than modern medicine. The method of conbined treatment of traditional Chinese medicine and western medicine can be effective in preventing fetal and neonatal hemolytic-induced complications, improve the quality of life of the newborn,is an ideal treatment antenatal maternal ABO hemolytic method, so it can be used in clinical application and has important significance to prevent the occurrence of the hemolytic disease of the newborn.ObjetiveBy research through the extracting free fetal DNA in the O-type blood plasma of pregnant women，using PCR-RFLP to detect fetal ABO blood group,we want to look for a fast, secure, high sensitivity and specificity new method of non-invasive prenatal diagnosis of Feto-Maternal ABO Blood Group Incompatibility;and give chinese and western medicine treatment options for these patients,so as to cure the Feto-Maternal AB0 Blood Group Incompatibility ,prevent and reduce morbidity and mortality of hemolytic disease of the newborn.MethodsWith the use of silicon membrane method , we extract fetal DNA from plasma of pregnant woman who is health and O-type blood, pregnant single-child and 13 to 37 weeks ,her husband is non-O blood type, detect fetal ABO blood group with PCR-RFLP method, amplify ABO glycosyltransferase genes in two specific fragments,afterward, use restriction enzyme Alu Ⅰ and Kpn Ⅰ to digest, enzyme electrophoresis determine fetal blood products. finally, determine blood type with the digestion products by electrophoresis . Blood test results by fetal diagnosis blood type incompatibility. Combined fetal blood type and maternal blood serum antibody titer of the test results , diagnose the Feto-Maternal Blood Group Incompatibility.Then treat the Feto-Maternal Blood Group Incompatibility patients using the method of combining chinese and western medicine.The prescriptions of traditional Chinese medicine is Yinchen Tuihuang soup, to consist ofartemisiae capillaries 12g、baical skullcap root 9g、prepared rhubarb 4.5g、licorice root 6g; The prescriptions of Western drugs is Vitamin C injection 3g joined the 10% in glucose solution 500ml, intravenous drip,or 50% in glucose solution 40ml add Vitamin C 500mg intravenous bolus, 1 time per day； oral vitamin E 100mg，1 time per day；at the same time the daily oxygen 2 times, 30min, a total of 10 days.ResultIn the 50 maternal plasma cases,detected 32 cases of fetal blood group, Where fetal blood type contain A-12 cases, B-11 cases, O-9 cases,check out the rate of 64%, born serological diagnosis confirmed 30 cases correctly, exact rate of 94%.The study indicate that in maternal blood existence Fetal DNA, we can use silicone membrane methods will be.The PCR-RFLP method detect of fetal ABO blood group is an effective non-invasive method, should be widely applied; Take advantage of free Fetal DNA can prenatal diagnosis of fetal ABO blood groups, simple and no injury,has important meaning of diagnosis and prevention of HDN.Following the treatment of combination therapy in chinese and western medicine, in 23 patients ,there are 20 cases of markedly effective treatment and 3 cases of effective treatment.The total effective rate is 100%. This description, the scheme is a way to reduce serum antibody titer, treatment of mothers and infants ABO, prevention and reduction of the hemolytic disease of the newborn morbidity and mortality.ConclusionUsed the free fetal DNA in maternal plasma, can detect fetal ABO blood, prenatal diagnosis the Feto-Maternal AB0 Blood Group Incompatibility. Infer from this, we can guass, the free Fetal DNA from blood of pregnant women can be helpful to prenatal diagnosis for a wide variety of fetal diseases and pathological pregnancies, it will be possible to replace the traditional prenatal diagnosis method, becoming a new noninvasive prenatal diagnosis, and applied to clinical.The efficacy which prevent and cure the Feto-Maternal AB0 Blood Group Incompatibility is satisfying ,by using the method of combining Chinese and Western medicine. The method is worthy to study in-depth and is used in clinical treatment.Key wordsFetal DNA; ABO blood group; The Feto-Maternal AB0 Blood Group Incompatibility; The treatment of Combining Chinese and Western medicineAuthor: LIU NaTutor: JING Ru-Cao陕西中医学院学位论文独创性声明及使用授权声明学位论文独创性声明本人郑重声明：所呈交的学位论文，是个人在导师的指导下，独立进行研究工作所取得的成果，无抄袭及编造行为。