1 Non-Invasive prenatal diagnosis of multiple endocrine neoplasia type 2A using COLDPCR combined with HRM genotyping analysis from maternal serum
Hada C.Macher1,Maria A.Martinez-Broca2, Amalia Rubio-Calvo1, Cristina Leon-Garcia1, Manuel Conde-Sanchez1,Alzenira Costa1,Elena Navarro2,Juan M.Guerrero1*
∥1.Department of Clinical Biochemistry,The Virgen del Roc?′o University Hospital(IBiS/CSIC/SAS/University of Seville),Seville,Spain
2.Department of Endocrinology,The Virgen del Roc?′o University Hospital(IBiS/CSIC/SAS/University of Seville),Seville,SpainPLoS One.2012,7(12):e51024.DOI:10.1371/journal.pone.0051024.Epub 2012 Dec 7.
多發(fā)性?xún)?nèi)分泌腺瘤II型(MEN2A)是常染色體顯性遺傳的單基因遺傳疾病,突變攜帶者多發(fā)甲狀腺髓樣癌。MEN2A是由于位于10q11.2的酪氨酸激酶c-RET原癌基因突變引起的。作者使用高分辨率溶解曲線(xiàn)分析法(HRM)對(duì)25例患者外周血DNA進(jìn)行驗(yàn)證性測(cè)試C634Y致病突變,準(zhǔn)確率達(dá)到100%。隨后對(duì)母體循環(huán)中游離胎兒DNA的測(cè)試表明,測(cè)試結(jié)果與PCR擴(kuò)增產(chǎn)物測(cè)序結(jié)果相符。HRM具有快速、無(wú)須對(duì)PCR產(chǎn)物進(jìn)一步處理的優(yōu)點(diǎn),可用于增加PCR檢測(cè)的敏感性。本文建立了用HRM對(duì)多發(fā)性?xún)?nèi)分泌腺瘤II型無(wú)創(chuàng)產(chǎn)前檢測(cè)方法,主要針對(duì)C634Y突變位點(diǎn)的檢測(cè)。
2 Rapid screening for chromosomal aneuploidies using array-MLPA Jing-Bin Yan1,2,Miao Xu1,Can Xiong1,Da-Wen Zhou1, Zhao-Rui Ren1, Ying Huang1,2, Monique Mommersteeg3,Rinie van Beuningen3,Ying-Tai Wang4,Shi-Xiu Liao4,F(xiàn)anyi Zeng1,2,5,Ying Wu1,3*and Yi-Tao Zeng1,2*
∥1.Institute of Medical Genetics,Children’s Hospital of Shanghai,Shanghai Jiao Tong University,Shanghai,P.R.China
2.Key Lab of Embryo Molecular Biology,Ministry of Health,and Shanghai Lab of Embryo and Reproduction Engineering,Shanghai,P.R.China
3.PamGene International BV, ‘s-Hertogenbosch, The Netherlands.4,Medical Genetic Institute of Henan Province,the People’s Hospital of Henan Province,Zhengzhou,P.R.China.5,Institute of Medical Sciences,Shanghai Jiao Tong University School of Medicine,Shanghai,P.R.China.
BMC Medical Genetics,2011 12:68.DOI:10.1186/1471-2350-12-68
多重連接依賴(lài)的探針擴(kuò)增技術(shù)(MLPA)能有效增補(bǔ)各種產(chǎn)前診斷技術(shù)的不足,但該技術(shù)最大局限是對(duì)連接產(chǎn)物大小的依賴(lài)性太強(qiáng),每次只能使用不到50對(duì)探針?;谶@些原因,作者進(jìn)一步研究了芯片多重連接依賴(lài)的探針擴(kuò)增技術(shù),首發(fā)芯片針對(duì)Dunchene肌營(yíng)養(yǎng)不良患者。該芯片對(duì)每個(gè)基因的3’和5’端各設(shè)計(jì)了一個(gè)探針,正常對(duì)照樣本的探針信號(hào)89.4%落在0.85~1.15,相對(duì)其他MLPA技術(shù)正常樣本信號(hào)區(qū)間0.7~1.3,區(qū)域更小。對(duì)161例外周血樣本、12例羊水樣本的檢測(cè)表明,有97.7%的樣本(其中包括所有羊水樣本)芯片多重連接依賴(lài)的探針擴(kuò)增技術(shù)與核型分析結(jié)果吻合。該研究表明,芯片多重連接依賴(lài)的探針擴(kuò)增技術(shù)可用于核型分析診斷的有效輔助手段。
3 Preliminary experience with cardiovascular magnetic resonance in evaluation of fetal cardiovascular anomalies
Su-Zhen Dong1,Ming Zhu1and Fen Li2*
∥1.Department of Radiology,Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine,Shanghai 200127,China
2.Department of Cardiology,Shanghai Children’s Medical Center, Shanghai Jiaotong University School of Medicine,Shanghai 200127,ChinaJournal of Cardiovascular Magnetic Resonance,2013,15:40
心血管畸形是最常見(jiàn)的先天性胎兒畸形。作者于2006年1月至2011年6月累積了68例懷有心血管畸形胎兒的孕婦,并對(duì)這些病例進(jìn)行常規(guī)產(chǎn)科超聲、超聲心動(dòng)圖和胎兒心血管磁共振檢測(cè)。將這些檢測(cè)結(jié)果與出生后檢查對(duì)比后發(fā)現(xiàn),79%(54/68)患者的胎兒心血管磁共振檢測(cè)結(jié)果與產(chǎn)后檢查結(jié)果一致。常規(guī)產(chǎn)科超聲結(jié)果與出生后檢查結(jié)果僅有46%(31/68)的一致性。由心血管專(zhuān)家進(jìn)行的超聲心動(dòng)圖檢測(cè)結(jié)果與產(chǎn)后檢查結(jié)果一致率為82%(56/68)。另外,有2%患者超聲心動(dòng)圖和磁共振皆誤診,15%患者被超聲心動(dòng)圖誤診但被磁共振確診,18%患者被磁共振誤診但被超聲心動(dòng)圖確診。被超聲心動(dòng)圖誤診但被磁共振確診的病例有無(wú)脾綜合征、多脾綜合征伴下腔靜脈中斷、三尖瓣關(guān)閉不全、右心室雙出口、雙主動(dòng)脈弓、右肺動(dòng)脈發(fā)育不全、法洛四聯(lián)癥以及左心發(fā)育不全綜合征。該研究表明胎兒心血管磁共振檢測(cè)可在超聲心動(dòng)圖檢測(cè)困難時(shí)進(jìn)行有效輔助。
4 Fetal-specific DNA methylation ratio permits noninvasive prenatal diagnosis of trisomy 21.
Elisavet A Papageorgiou1, Alex Karagrigoriou2,Evdokia Tsaliki1,3,4,Voula Velissariou3,Nigel P Carter5&Philippos C Patsalis1
∥1.Cytogenetics and Genomics Department,The Cyprus Institute of Neurology and Genetics,Nicosia,Cyprus
2.Department of Mathematics and Statistics,University of Cyprus,Nicosia,Cyprus
3.Department of Cytogenetics and Molecular Biology, Mitera Hospital,Athens, Greece.4Faculty of Biology,School of Sciences,National and Kapodistrian University of Athens,Athens,Greece.5,Molecular Cytogenetics Department,The Wellcome Trust Sanger Institute,Cambridge,Hinxton,UKNat Med,2011,17(4):510-513
作者在篩查40例正常胎兒和21三體胎兒母外周血游離DNA樣本已知高堿基化區(qū)域HYP113和低堿基化區(qū)域 U122后發(fā)現(xiàn),21三體胎兒差異堿基化區(qū)域比值常大于1,正常胎兒比值常小于1。作者利用這一發(fā)現(xiàn),開(kāi)發(fā)甲基化DNA免疫共沉淀技術(shù)(MeDIP)合并實(shí)時(shí)定量PCR進(jìn)行21三體無(wú)創(chuàng)產(chǎn)前檢查。40例盲篩結(jié)果顯示該檢測(cè)方法準(zhǔn)確率100%,為無(wú)創(chuàng)產(chǎn)前檢查提供了新方法和技術(shù)。
5 MeDIP real-time qPCR of maternal peripheral blood reliably identifies trisomy 21
Evdokia Tsaliki1,2,3, Elisavet A.Papageorgiou1,2, Christiana Spyrou1,George Koumbaris1,2,Elena Kypri1,Skevi Kyriakou2,Chrysovalanto Sotiriou1,Evi Touvana1, Anna Keravnou1, Alex Karagrigoriou4,Klea Lamnissou3,Voula Velissariou5and Philippos C.Patsalis2*
∥1.NIPD Genetics Ltd,Nicosia,Cyprus
2.The Cyprus Institute of Neurology and Genetics,Nicosia,Cyprus
3.Department of Genetics and Biotechnology,F(xiàn)aculty of Biology,School of Science,National and Kapodistrian University of Athens,Panepistimiopolis,Ilissia,Athens,Greece
4.Department of Mathematics and Statistics,University of Cyprus,Nicosia,Cyprus
5.Department of Genetics and Molecular Biology,Mitera Hospital,Athens,GreecePrenatal Diagnosis,2012,32:996-1001
作者于2011年開(kāi)發(fā)使用12組差異堿基化區(qū)域進(jìn)行甲基化DNA免疫共沉淀技術(shù)無(wú)創(chuàng)產(chǎn)前檢查。本文中作者使用75例已知診斷結(jié)果的母親外周血胎兒游離DNA樣本,100例已檢測(cè)但雙盲樣本對(duì)前述診斷方法進(jìn)行驗(yàn)證。驗(yàn)證結(jié)果顯示該方法特異性99.2%,敏感性100%,提示這一檢測(cè)手段對(duì)檢測(cè)21三體無(wú)創(chuàng)檢測(cè)非常有效。作者將進(jìn)一步使用更大樣本量(700~1000例)對(duì)此方法進(jìn)行驗(yàn)證,并通過(guò)研究簡(jiǎn)化這一檢測(cè)方法的實(shí)驗(yàn)步驟。
6 Early fetal gender determination using realtime PCR analysis of cell-free fetal DNA during 6th-10th weeks of gestation.Khorram Khorshid HR, Zargari M,Sadeghi MR,Edallatkhah H,Shahhosseiny MH,Kamali K.
∥Genetic Research Centre,University of Social Welfare and Rehabilitation Sciences,Tehran,Iran.
Acta Med Iran,2013 ,51(4):209-214.
母親外周血胎兒游離DNA樣本相對(duì)于傳統(tǒng)產(chǎn)前診斷檢查有著高效、低風(fēng)險(xiǎn)的優(yōu)勢(shì)。無(wú)創(chuàng)產(chǎn)前檢查的一個(gè)重要應(yīng)用方向是出生前性別檢測(cè),以對(duì)可能患有性別相關(guān)的遺傳病胎兒做出判斷。作者抽取了80例6~10孕周孕婦外周血樣本,采用實(shí)時(shí)定量PCR檢測(cè)了SRY、DYS14、DAZ基因序列。檢測(cè)結(jié)果同出生后實(shí)際結(jié)果進(jìn)行比對(duì),得到97.3%的特異性和97.3%的敏感性指標(biāo)。由此作者得出結(jié)論,利用實(shí)時(shí)定量PCR對(duì)母親外周血胎兒游離DNA樣本進(jìn)行檢測(cè)可盡早對(duì)胎兒性別做出判斷,有利于患兒父母及早決定是否終止妊娠。
7 Diagnostic accuracy of ultrasound and MRI in the prenatal diagnosis of placenta accreta.
Maher MA,Abdelaziz A,Bazeed MF.
∥Armed Forces Hospital Southern Region,Khamis Mushyat,Saudi Arabia;Obstetrics and Gynaecology Department,F(xiàn)aculty of Medicine, Menofiya University, Shebin Elkom,Egypt.
Acta Obstet Gynecol Scand,2013 May 28.DOI:10.1111/aogs.12187.
此文章是沙特阿拉伯地區(qū)對(duì)胎盤(pán)植入和低位胎盤(pán)進(jìn)行超聲和磁共振比較的前瞻性分析研究。對(duì)577例患者研究表明,超聲正確診斷了39例胎盤(pán)植入中的33例,排除514例非胎盤(pán)植入中的512例,敏感性95.1%,特異性95.5%。磁共振檢測(cè)成功診斷了20例胎盤(pán)植入中的6例,排除了20例非胎盤(pán)植入中的10例,敏感性85.7%,特異性76.9%。作者得出結(jié)論,超聲能有效診斷胎盤(pán)植入病例,如遇到疑難病例,可由磁共振進(jìn)一步輔助診斷。
8 Outcome after second-trimester amniocentesis and first-trimester chorionic villus sampling for prenatal diagnosis in multiple gestations.
Enzensberger C,Pulvermacher C,Degenhardt J,Kawecki A,Germer U,Weichert J,Krapp M,Gembruch U,Axt-Fliedner R.
∥OB&GYN,Division of Prenatal Medicine,Justus-Liebig-University,Gie?en.
Ultraschall Med.2013 May 21.
作者隨訪了288例多胎妊娠孕婦(總計(jì)637例胎兒)在羊水穿刺、絨毛膜取樣2周后意外流產(chǎn)(無(wú)染色體異常、無(wú)產(chǎn)科并發(fā)癥)情況。排除染色體異常、產(chǎn)科并發(fā)癥后,病例中共計(jì)132例羊水穿刺、44例絨毛膜取樣??偱咛チ魇蕿?%(14/176),其中羊水穿刺妊娠終止率為6.1%(n=8),絨毛膜取樣妊娠終止率為13.6%(n=6)。這一比率與其他文獻(xiàn)報(bào)道相符,作者提出結(jié)論,在對(duì)多胎妊娠孕婦進(jìn)行相關(guān)檢測(cè)前,應(yīng)告知檢測(cè)引起的妊娠終止風(fēng)險(xiǎn)。
9 Multiplex ligation-dependent probe amplification(MLPA):a reliable alternative for fetal chromosome analysis?
Chitty LS,Kistler J,Akolekar R,Liddle S,Nicolaides K,Levett L.
∥University College London Institute of Child Health and Fetal Medicine Unit,UCLH NHS Foundation Trust,London,UK
J Matern Fetal Neonatal Med,2012,25(8):1383-1386.
作者使用多重連接依賴(lài)的探針擴(kuò)增技術(shù)(MLPA)檢測(cè)了476例核型分析后的絨毛膜或羊水樣本。多重連接依賴(lài)的探針擴(kuò)增技術(shù)、核型分析以及定量熒光PCR檢測(cè)方法共檢出了190例異常,其中124例三體征、21例性染色體相關(guān)異常、14例三倍體以及31例染色體重組或嵌合體。其中三體征都能被所有方法檢出,三倍體只能被核型分析以及定量熒光PCR檢測(cè)方法檢出。在19例高風(fēng)險(xiǎn)重組和嵌合體病例,染色體核型分析檢出13例,MLPA檢出18例,定量熒光PCR未能檢出病例。故而作者得出結(jié)論,使用特異性探針的MLPA較傳統(tǒng)遺傳學(xué)分析更能檢出高風(fēng)險(xiǎn)病例,將MLPA和定量熒光PCR相結(jié)合能提高診斷效率。