多重荧光定量PCR方法检测X-连锁鱼鳞病家系女性携带者

Multiplex quantitative PCR detection for female carrier in an X-linked ichthyosis family

目的分析一个汉族X-连锁鱼鳞病家系中类固醇硫酸酯酶（STS）基因的致病突变，明确家系中3名女性可能携带者身份，并比较不同方法在STS基因缺失突变检测中的应用。方法采用普通PCR方法，对家系中男性先证者进行STS基因10个外显子的扩增，明确是否存在缺失。采用多重荧光定量聚合酶链反应（QF-PCR）方法对X-连锁鱼鳞病家系部分女性成员进行STS基因定量分析。并应用荧光原位杂交（FISH）技术进行验证。结果普通PCR方法检测到先证者STS基因第1—10号外显子缺失；多重QF-PCR方法检测到先证者母亲为STS基因缺失突变携带者，先证者妹妹及表妹均不存在该缺失。FISH检测结果与QF-PCR结果相同。结论对于STS基因完全缺失型患者及女性携带者，多重QF—PCR和FISH两种方法均能有效检测，但多重QF—PCR方法对STS基因缺失的检测操作方便、自动化程度高、检测通量高。

Objective To analyze the pathogenic mutation of an X-linked ichthyosis （XLI） family, and identify the genetic diagnosis of three probable female carriers in this family. To evaluate the availability of different detect methods for steroid sulfatase （STS） gene mutation. Methods Peripheral blood samples were collected from the family, including the proband, proband＇s mother, younger sister, and younger female cousin, and 10 males and 10 females as controls. Ordinary PCR was used to detect whether there was STS gene deletion in the male proband. Then, multiplex quantitative fluorescent PCR （QF-PCR） was used to detect the STS gene in the proband and his 3 female family members. Fluorescence in situ hybridization （FISH） was used to authenticate the results of multiplex QF-PCR method. Results No amplified product of the exons 1 - 10 of STS gene deletion was detected by ordinary PCR in the proband. The proband＇s mother was diagnosed as a carrier, but his sister and cousin were diagnosed as normal females by multiplex QF-PCR. FISH confirmed the results of multiplex QF-PCR. Conclusion Both multiplex QF-PCR and FISH are effective to detect the complete deletion mutation of STS gene and identify the female carrier, and multiplex QF-PCR is more convenient and automatic compared with FISH.