Funari Mariana F A, Jorge Alexander A L, Souza Silvia C A L, Billerbeck Ana E C, Arnhold Ivo J P, Mendonca Berenice B, Nishi Mirian Y
Unidade de Endocrinologia do Desenvolvimento, Hospital das Clinicas, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brazil.
Eur J Med Genet. 2010 Sep-Oct;53(5):234-8. doi: 10.1016/j.ejmg.2010.06.001. Epub 2010 Jun 9.
SHOX haploinsufficiency causes a wide spectrum of short stature phenotypes, such as Leri-Weill dyschondrosteosis (LWD) and disproportionate short stature (DSS). SHOX deletions are responsible for approximately two thirds of isolated haploinsufficiency; therefore, it is important to determine the most appropriate methodology for detection of gene deletion. In this study, three methodologies for the detection of SHOX deletions were compared: the fluorescence in situ hybridization (FISH), microsatellite analysis and multiplex ligation-dependent probe amplification (MLPA). Forty-four patients (8 LWD and 36 DSS) were analyzed. The cosmid LLNOYCO3'M'34F5 was used as a probe for the FISH analysis and microsatellite analysis were performed using three intragenic microsatellite markers. MLPA was performed using commercial kits. Twelve patients (8 LWD and 4 DSS) had deletions in SHOX area detected by MLPA and 2 patients generated discordant results with the other methodologies. In the first case, the deletion was not detected by FISH. In the second case, both FISH and microsatellite analyses were unable to identify the intragenic deletion. In conclusion, MLPA was more sensitive, less expensive and less laborious; therefore, it should be used as the initial molecular method for the detection of SHOX gene deletion.
矮小同源框基因（SHOX）单倍剂量不足会导致多种身材矮小的表型，如莱里-韦伊软骨发育不全（LWD）和不成比例身材矮小（DSS）。SHOX基因缺失约占孤立性单倍剂量不足病例的三分之二；因此，确定检测基因缺失的最合适方法很重要。在本研究中，比较了三种检测SHOX基因缺失的方法：荧光原位杂交（FISH）、微卫星分析和多重连接依赖探针扩增（MLPA）。对44例患者（8例LWD和36例DSS）进行了分析。黏粒LLNOYCO3'M'34F5用作FISH分析的探针，使用三个基因内微卫星标记进行微卫星分析。MLPA使用商业试剂盒进行。12例患者（8例LWD和4例DSS）通过MLPA检测到SHOX区域存在缺失，2例患者与其他方法产生了不一致的结果。在第一个病例中，FISH未检测到缺失。在第二个病例中，FISH和微卫星分析均未能识别基因内缺失。总之，MLPA更敏感、成本更低且操作更简便；因此，它应作为检测SHOX基因缺失的初始分子方法。
