焦磷酸解激活聚合反应在McCune-Albright综合征基因突变检测中的应用

doi:10.7504/ek2014080609

摘要/Abstract

摘要：

目的应用新一代核酸扩增技术“焦磷酸解激活聚合反应（PAP）”检测McCune-Albright综合征（MAS）致病基因GNAS1的已知热点突变,并探讨其在MAS诊断中的应用价值。方法依据临床特征将2006年1月至2013年9月上海交通大学医学院附属瑞金医院儿内科36例疑诊MAS患儿分为典型组（非GnRH依赖型性早熟和骨纤维结构发育不良，伴或不伴皮肤牛奶咖啡斑）与非典型组（非GnRH依赖型性早熟，伴或不伴皮肤牛奶咖啡斑）；33例健康成年志愿者作为对照组。针对MAS致病基因（GNAS1）的已知突变热点（R201H/C/L），分别采用实时荧光PAP技术与普通PCR技术进行检测比较。结果在36例临床疑诊MAS患儿中，经实时荧光PAP技术发现9例（25%）存在GNAS1已知突变（4例R201H、5例R201C和0例R201L），其中典型组突变发现率为63.64%（7/11），非典型组为8.00%（2/25）；对照组均未发现已知突变。而由普通PCR技术亦未发现已知突变（检出率0）。实时荧光PAP技术的已知突变检测率明显高于普通PCR技术（P＜0.05）。结论我国MAS患者存在GNAS1基因R201H和R201C突变热点，PAP技术可有助于临床对MAS的分子病理诊断。

关键词: 焦磷酸解激活聚合反应, McCune-Albright 综合征, 分子诊断

Abstract:

Abstracts： Objective To detect McCune-Albright syndrome （MAS） causing gene GNAS1 mutation hotspots by a new generation nucleic acid amplification “pyrophosphorolysis activated polymerization （PAP）” and to explore its value in the diagnosis of MAS. Methods Thirty-six children clinically verified as MAS were enrolled. They were subdivided into typical groups （with GnRH independent precocious puberty and polyostotic fibrous dysplasia，with or without skin cafē au lait spots） and atypical groups （with GnRH independent precocious puberty ,with or without skin cafē au lait spots） according to their phenotypes. In addition，a cohort of thirty-three healthy adults were enrolled as control group. Two techniques （real-time fluorescence PAP and common PCR） were respectively used to detect GNAS1 mutation hotspots and were compared. Results By the technique of real-time fluorescence PAP，two kinds of mutations were identified in nine of thirty-six MAS patients （4 with R201H and 5 with R201C，none with R201L），the positive findings being 9/36 （25%） while none by the common PCR. The mutation rates were significantly higher in the typical group as 63.64%（7 out of 11 cases）but 8%（2/25）in atypical group and all negative in control group. The technique of real-time fluorescence PAP revealed dominant priority in efficiency of molecular diagnosis of MAS compared with the common PCR. Conclusion GNAS1 gene mutation hotspots （R201H and R201C） are also present in children with MAS，and PAP technology can contribute to clinical molecular diagnosis of MAS.

Key words: pyrophosphorolysis activated polymerization（PAP）, McCune-Albright syndrome（MAS）, molecular diagnosis

中图分类号:

Ｒ72

秦雪艳，王伟，董治亚，陆文丽，倪继红，肖园，王德芬. 焦磷酸解激活聚合反应在McCune-Albright综合征基因突变检测中的应用[J]. 中国实用儿科杂志, DOI: 10.7504/ek2014080609.

QIN Xue-yan，WANG Wei，DONG Zhi-ya，LU Wen-li，NI Ji-hong，XIAO Yuan，WANG De-fen.. Application of pyrophosphorolysis activated polymerization in detecting gene mutation of McCune-Albright syndrome.[J]. Acta Metallurgica Sinica, DOI: 10.7504/ek2014080609.

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