摘要
目的分析应用多重连接依赖探针扩增技术(Multiplex ligation dependent probe amplification,MLPA)和免疫荧光染色技术来诊断单基因遗传病假肥大型肌营养不良症(DMD/BMD)的临床价值。方法运用多重连接依赖探针扩增技术对150例(120例DMD,30例BMD)患者的dystrophin基因的缺失或是重复突变进行筛查,同时运用免疫荧光染色技术检测150例假肥大型肌营养不良症患者的肌组织中抗肌营养不良蛋白表达,同时关注蛋白定位情况,以2例正常人的肌组织作为对照。结果多重连接依赖探针扩增技术检测150例患者中92例患者有外显子缺失,9例有外显子重复,免疫荧光染色技术检测证实了对照组抗肌营养不良蛋白定位在肌细胞膜上,且染色阳性,DMD患者肌膜完全无显色,BMD患者染色弱阳性,荧光显微镜下见沿着肌细胞膜分布的间断的且呈斑片状分布的荧光带。结论150例DMD/BMD患儿经MLPA技术分析,101例由缺失或是重复突变所致,也能推断抗肌营养不良蛋白缺乏或表达异常是DMD/BMD基本病理基础,因此运用此两种方法综合诊断有助于DMD和BMD的临床诊断和鉴别。
Objective:To explore the clinical value of Multiplex ligation dependent probe amplification combined with immunofluorescence in the detection of pseudohypertrophic muscular dystrophy.Methods:MLPA was used to screen the dystrophic gene deletion and repeat mutation in 150 patients(120 patients are DMD,30 patients are BMD),and needle aspiration biopsy was used to get the muscle tissue of 150 patients with pseudohypertrophic muscular dystrophy.Immunofluorescence staining was used to detect the expression and location of dystrophin,and twonormal human myocytes were used as control.Results:In 150 patients with MLPA,92 patients had exon deletion,9 patients had exon duplication.Immunofluorescenestaining confirmed that the control group had positive staining on the sarolemma,negative staining on the nucleus and cytoplasm,DMD patients had no staining on the sarcolemma,BMD patients had weak positive staining,and intermittent patchy fluorescent bands distributed along the sarcolemma could be seen.Conclusion:In 150 cases of DMD/BMD,101 cases of them were caused by deletion and repetitive mutation.Dystrophin deficiency or abnormal expression is the basic pathological basis of DMD/BMD.The comprehensive diagnosis of two methods is helpful for the diagnosis and discrimination of DMD/BMD.
作者
闫杨
王红梅
YAN Yang;WANG Hong-mei(Lab of Hematology,Air Force Hospital,northern theater,PLA,LiaoNing,Shenyang 110042)
出处
《中国优生与遗传杂志》
2020年第6期659-661,665,共4页
Chinese Journal of Birth Health & Heredity
