精子荧光原位杂交技术在胚胎植入前遗传学诊断中的作用

Application of sperm fluorescence in situ hybridization in preimplantation genetic diagnosis

目的初步探讨精子荧光原位杂交（FISH）技术在男性染色体异常胚胎植入前遗传学诊断（PGD）中的作用。方法2006年7月—2008年8月，选择于郑州大学第一附属医院生殖医学中心就诊的9对因男方染色体异常而导致不孕的夫妇，其中包括染色体罗氏易位携带者7例，相互易位携带者1例，克氏综合征患者1例。采用FISH技术对精子和胚胎进行分析，并分析精子和胚胎FISH分析结果之间的相关性。结果（1）9例患者的精子FISH分析共计数精子8568个，无荧光信号24个。7例罗氏易位携带者正常或平衡（即染色体平衡易位）精子比例为85．71％（6045／7053）；1例染色体相互易位携带者正常或平衡精子比例为30．42％（306／1006），1例克氏综合征患者正常精子比例为68．76％（350／509）。（2）共活检158个胚胎，成功固定135个，其中111个出现明显诊断信号，36个为染色体正常或平衡易位胚胎（26．7％），75个为染色体异常胚胎。共移植21个胚胎，成功妊娠1例，现已分娩健康婴儿。7例罗氏易位携带者中，正常或平衡胚胎比例为29．0％（31／107），1例相互易位携带者中正常或平衡胚胎比例为6．3％（1／16），1例克氏综合征患者正常胚胎比例为33．3％（4／12）。（3）PGD中正常精子的比例与正常胚胎的比例呈正相关关系（r=0．75，P=0．02）。结论精子FISH分析对PGD前生殖遗传咨询有重要的临床意义。

Objective To investigate the role of sperm fluorescence in situ hybridization （FISH） in preimplantation genetic diagnosis （PGD） for male chromosomal disorders. Methods From Jul. 2006 to Aug. 2008, FISH was performed in sperm and embryo of 9 infertile couples due to male chromosomal abnormality including 7 couples with Robertsonian translocation, one couple with reciprocal translocation and one couple with Klinefeher＇s syndrome. Correlation analysis was performed between sperm and embryo FISH results. Results （1） FISH analysis of 8568 sperms showed 24 sperms had no fluorescence signals. The rate of normal/balanced sperm of carriers were 85.71% （ 6045/7053 ） in seven Robertsonian translocation, 30. 42% （306/1006） in one reciprocal translocation and 68. 76% （350/509） in Klinefelter＇s syndrome. （2） A total of 158 embryos were biopsied, of which 135 embryos were successfully fixed for FISH. A hundred and one embryos exhibit informative signal including 36 normal/balanced embryos and 75 abnormal embryos. Twenty-one embryos were transferred and one couple obtained successful term pregnancy. The rate of normal/balanced embryo were 29. 0% （31/107） in 7 carriers of Robertsonian translocation, 6. 3% （1/16） in one reciprocal translocation and 33.3% （4/12） in Klinefelter＇s syndrome. （3） A positive correlated relationship was found between the percentage of normal embryo and the percentage of normal sperm （ r = 0. 75 ,P =0. 02）. Conclusion It is advisable to recommend the sperm FISH analysis for being routinely incorporated into the genetic screening offered prior to preimplantation genetic diagnosis.