本文選題：Prader-Willi + 綜合征��； 參考：《安徽醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的 Prader-Willi綜合征（PWS）的臨床診斷標(biāo)準(zhǔn)已于1993年由Holm等人提出，并得到公認(rèn)，但根據(jù)我院收集的資料發(fā)現(xiàn)，我國PWS患兒的臨床表型與Holm等總結(jié)的診斷標(biāo)準(zhǔn)存在差異，故本研究旨在分析我國PWS新生兒期的臨床表現(xiàn)，為臨床早期篩選及進(jìn)一步行分子診斷提供幫助。同時(shí)由于不同發(fā)病機(jī)制的Prader-Willi綜合征在臨床表現(xiàn)、預(yù)后和遺傳風(fēng)險(xiǎn)上均存在差異，而目前臨床常用確診方法MS-PCR不能區(qū)分發(fā)病機(jī)制，本研究采用甲基化特異性多重連接依賴的探針擴(kuò)增（MS-MLPA）技術(shù)和短串聯(lián)重復(fù)序列（STR）技術(shù)分析Prader-Willi綜合征的遺傳機(jī)理，分析MS-MLPA在診斷PWS及分辨發(fā)病機(jī)制上的臨床實(shí)用性。 方法 1.采用回顧性研究方法，將2009年4月至2011年8月收集的13例確診為PWS的新生兒的臨床表型作為研究資料，根據(jù)Holm等所提臨床診斷標(biāo)準(zhǔn)中的主、次要標(biāo)準(zhǔn)分別記錄每個(gè)新生兒的表型存在與缺失情況，進(jìn)而總結(jié)分析我國PWS新生兒期典型臨床特征。 2.取臨床進(jìn)行甲基化特異性PCR確診的16例PWS患兒的外周血樣本，重新提取DNA，采用甲基化特異性多重連接依賴的探針擴(kuò)增技術(shù)（MS-MLPA）試劑盒Me028進(jìn)行基因檢測分析。同時(shí)取其中收集到的6例PWS患兒及其父母外周血提取的DNA，采用短串聯(lián)重復(fù)序列（STR）技術(shù)進(jìn)行基因檢測分析。 結(jié)果 1.新生兒期主要表現(xiàn)有皮膚色素減退13例（100%，該點(diǎn)明顯高于國外報(bào)道），中樞性肌張力低下13例（100%，輕重程度不同）,性腺發(fā)育不良12例（92.31%），喂養(yǎng)困難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患兒孕母高齡9例（69.23%），羊水污染8例（61.54%），羊水過多3例（23.08%），胎膜早破5例（38.46%）,異常胎位4例（30.78%），宮內(nèi)窘迫9例（69.23%），，是圍生期母嬰常見并發(fā)癥。 2. MS-MLPA檢測結(jié)果與甲基化特異性PCR檢測結(jié)果一致，16例患兒均為PWS陽性患兒，且檢測出16例PWS病例中4例源于母源性同源二倍體，12例源于父源性15q11-q13區(qū)域缺失。并經(jīng)STR證實(shí)了MS-MLPA的基因來源分析的準(zhǔn)確性。 結(jié)論 1.新生兒期皮膚色素減退及中樞性肌張力低下可作為我國新生兒進(jìn)一步行PWS分子診斷的初步篩選標(biāo)準(zhǔn)。 2. MS-MLPA能夠確診PWS，并且能將PWS父源性缺失型和母源性同源二倍體兩種主要發(fā)病機(jī)制鑒別開的一種快捷、可靠的實(shí)驗(yàn)診斷技術(shù)。
[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.
【學(xué)位授予單位】：安徽醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R725.8

【參考文獻(xiàn)】

相關(guān)期刊論文 前2條

1 王薇;吳曉燕;宋紅梅;邱正慶;魏珉;;甲基化特異性PCR方法診斷Prader-Willi綜合征[J];中國當(dāng)代兒科雜志;2008年04期

2 朱麗娜;何璽玉;王春枝;楊曉;劉欣;王蔚;馬寧;劉海洪;王艷;封志純;;Prader-Willi綜合征的分子遺傳學(xué)診斷與機(jī)制研究[J];山西醫(yī)科大學(xué)學(xué)報(bào);2008年12期

本文編號：2012453