Prader-Willi综合征临床表型分析与分子诊断学研究

发布时间：2018-06-13 03:16

本文选题：Prader-Willi + 综合征　；参考：《安徽医科大学》2012年硕士论文

【摘要】：目的 Prader-Willi综合征（PWS）的临床诊断标准已于1993年由Holm等人提出，并得到公认，但根据我院收集的资料发现，我国PWS患儿的临床表型与Holm等总结的诊断标准存在差异，故本研究旨在分析我国PWS新生儿期的临床表现，为临床早期筛选及进一步行分子诊断提供帮助。同时由于不同发病机制的Prader-Willi综合征在临床表现、预后和遗传风险上均存在差异，而目前临床常用确诊方法MS-PCR不能区分发病机制，本研究采用甲基化特异性多重连接依赖的探针扩增（MS-MLPA）技术和短串联重复序列（STR）技术分析Prader-Willi综合征的遗传机理，分析MS-MLPA在诊断PWS及分辨发病机制上的临床实用性。方法 1.采用回顾性研究方法，将2009年4月至2011年8月收集的13例确诊为PWS的新生儿的临床表型作为研究资料，根据Holm等所提临床诊断标准中的主、次要标准分别记录每个新生儿的表型存在与缺失情况，进而总结分析我国PWS新生儿期典型临床特征。 2.取临床进行甲基化特异性PCR确诊的16例PWS患儿的外周血样本，重新提取DNA，采用甲基化特异性多重连接依赖的探针扩增技术（MS-MLPA）试剂盒Me028进行基因检测分析。同时取其中收集到的6例PWS患儿及其父母外周血提取的DNA，采用短串联重复序列（STR）技术进行基因检测分析。结果 1.新生儿期主要表现有皮肤色素减退13例（100%，该点明显高于国外报道），中枢性肌张力低下13例（100%，轻重程度不同）,性腺发育不良12例（92.31%），喂养困难11例（84.62%）；其他表型有特殊面容5例（38.46%），唾液粘稠5例（38.46%）。此外PWS患儿孕母高龄9例（69.23%），羊水污染8例（61.54%），羊水过多3例（23.08%），胎膜早破5例（38.46%）,异常胎位4例（30.78%），宫内窘迫9例（69.23%），，是围生期母婴常见并发症。 2. MS-MLPA检测结果与甲基化特异性PCR检测结果一致，16例患儿均为PWS阳性患儿，且检测出16例PWS病例中4例源于母源性同源二倍体，12例源于父源性15q11-q13区域缺失。并经STR证实了MS-MLPA的基因来源分析的准确性。结论 1.新生儿期皮肤色素减退及中枢性肌张力低下可作为我国新生儿进一步行PWS分子诊断的初步筛选标准。 2. MS-MLPA能够确诊PWS，并且能将PWS父源性缺失型和母源性同源二倍体两种主要发病机制鉴别开的一种快捷、可靠的实验诊断技术。
[Abstract]:Objective the clinical diagnostic criteria of Prader-Willi syndrome PWSs were put forward by Holm et al in 1993, and were generally accepted. However, according to the data collected in our hospital, the clinical phenotype of children with PWS in China is different from that of Holm et al. The purpose of this study is to analyze the clinical manifestations of PWS in neonates in China, and to provide help for early screening and further molecular diagnosis of PWS. At the same time, the clinical manifestations, prognosis and genetic risk of Prader-Willi syndrome with different pathogenesis were different. However, MS-PCR, a commonly used clinical diagnosis method, could not distinguish the pathogenesis of Prader-Willi syndrome. In this study, we analyzed the genetic mechanism of Prader-Willi syndrome by using methylation specific multiple connection-dependent probe amplification (MS-MLPA) technique and short tandem repeat (STR) technique, and analyzed the clinical usefulness of MS-MLPA in diagnosing PWS and distinguishing pathogenesis. Method 1. The clinical phenotypes of 13 newborns diagnosed as PWS from April 2009 to August 2011 were used as the data of retrospective study, according to Holm et al. The secondary criteria recorded the phenotypic status of each newborn and analyzed the typical clinical characteristics of PWS in China. 2. The peripheral blood samples of 16 PWS patients confirmed by methylation specific polymerase chain reaction (PCR) were collected and the DNA was reextracted. The MS-MLPA kit Me028 was used to detect and analyze the DNA. The DNA extracted from peripheral blood of 6 children with PWS and their parents were analyzed by short tandem repeat (STR) technique. Result 1. There were 13 cases of hypodermic skin pigmentation in neonatal stage, which was obviously higher than that reported abroad, 13 cases of central hypotonia with different degrees of severity, 12 cases of gonadal dysplasia, and 11 cases of feeding difficulties (84.62%). Other phenotypes included 5 special faces (38.46) and salivary thickening (38.46). In addition, 9 elderly women with PWS, 61.54 with amniotic fluid contamination, 23.08 with amniotic fluid, 5 with premature rupture of membranes, 4 with abnormal fetal position and 69.23 with intrauterine distress were common complications in perinatal period. The results of MS-MLPA and methylation-specific PCR were consistent with that of methylation-specific polymerase chain reaction in 16 cases. Among 16 cases, 4 cases were from maternal autodiploid and 12 cases from paternal 15q11-q13 region deletion. The accuracy of gene source analysis of MS-MLPA was confirmed by STR. Conclusion 1. Hypodermic skin and central dystonia can be used as screening criteria for further molecular diagnosis of PWS in neonates in China. 2. MS-MLPA can be used to diagnose PWSs, and it is a fast and reliable experimental diagnostic technique to distinguish the two main pathogenesis of PWS parenchymal deletion and maternal autodiploid.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R725.8

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