多重连接依赖探针扩增技术在稽留流产绒毛组织染色体核型分析中的应用

Karyotype analysis of chorionic villi from pregnant women with missed abortion using multiplex ligation-dependent probe amplification

目的探讨多重连接依赖探针扩增（MLPA）技术在稽留流产绒毛组织染色体核型分析中的应用。方法选择2008年2-10月于深圳市妇幼保健院就诊，经激素水平测定、B超和临床检查确诊为稽留流产患者91例为病例组；随机抽样方法选择同期20例经激素水平测定、B超和临床检查为正常妊娠，要求人工流产者为对照组。人工流产术中无菌条件下获取两组妇女的绒毛组织，培养后每份样本分别采用传统细胞遗传学G显带染色体核型分析方法、同时提取DNA采用MLPA技术分析染色体异常，并与传统染色体核型分析结果进行比较。结果91例病例组样本中，有84例（92％）采用MLPA技术进行染色体核型分析的非整倍体结果与传统染色体核型分析方法的结果一致，其中包括正常核型40例、常染色体三体29例、常染色体双三体1例、X染色体单体合并常染色体三体1例、X染色体单体10例、嵌合性X染色体单体2例和1例结构异常46，XX，der（5）t（5；8）（p1．2；q1．2）；其余结果不一致的7例中，采用传统染色体核型分析方法检测出2例三倍体和5例四倍体，而采用MLPA技术检测结果均为正常的二倍体。20例对照组样本两种技术的分析结果均一致。结论MI，PA技术分析染色体非整倍体异常是一种简单、快速且有效的方法，具有临床实际应用价值。

Objective To evaluate the clinical value of multiplex ligation-dependent probe amplification （MLPA） technique used in karyotype analysis of chorionic villi from missed abortion. Methods Feb 2008 to Oct 2008, 91 patients with missed abortion diagnosed by hormonal measurement, type B ultrasound and physical exam matched with 20 normal pregnant women undergoing artificial abortion were enrolled in this study. Chorionic villi was obtained by suction dilation and curettage in aseptic condition, then those villi was cultured and analyzed by traditional cytogenetic karyotyping method, in the mean time, the DNA extracted from villi was detected by MLPA. The results of chromosomal G-banding of chorionic villi were compared between two methods. Results The diagnostic concordance of MLPA and traditional karyotyping was observed in 92% （84/91） cases, there were 84 cases in the case group with diagnostic concordance by traditional karyotyping and MLPA except 7 cases of euploidy could not be detected by MLPA. The 84 cases included 40 normal karyotype, 29 trisomy of euchromosome, 1 double trisomy of euehromosome, 10 monosomy X , 1 monosomy X combined with trisomy of euehromosome, 2 chimaera of X chromosome, 1 structural abnormity of euehromosome. Among 7 cases with discordance diagnosis, 2 cases with trisomy and 5 cases with tetrasomy of euchromosome were identified in traditional karyotyping, however, they were all diagnosed with normal disomy by MLPA. Of 20 villi from normal pregnancy, two methods got the consistent results. Conclusion The MLPA was rapid and efficacy method used for analyzing aneuploids in ehorionic villi.