摘要目的:基于多重PCR方法针对Turner综合征患者，建立快速检测Y染色体特有序列的多重PCR方法。方法:在Y染色体不同区域选择9个基因设计引物，其中Y染色体短臂特有基因为 SRY、TBL1Y、TSPY，长臂特有基因为 DDX3Y、HSFY1、RPS4Y2、CDY1，以及X和Y染色体拟常染色体区(PAR)短臂的 SHOX和长臂的 SPRY3基因。建立并优化Y染色体9个基因的多重PCR方法，通过使用基因组DNA不同用量对多重PCR进行灵敏性测试，并对确诊的36例Turner综合征患者(2例患者核型携带mar，1例患者核型为45，X[40]/47XYY[21])以及1例核型为46，X，+mar的男性矮小症患者进行检测。 结果:优化多重PCR反应体系(50 μL)结果，每对引物上下游的终浓度为0.1 μM时，9对引物的多重PCR反应扩增出与理论大小一致的目的条带，无非特异性扩增，条带清晰易区分。在灵敏性试验中，多重PCR反应体系的基因组DNA用量低至1 ng时条带仍清晰可见。使用该方法检测36例Turner综合征患者，1例核型为45，X[40]/47XYY[21]的Turner综合征患者扩增出Y染色体特有目的基因，35例Turner综合征患者只扩增出 SHOX和 SPRY3两个目的基因，而无Y染色体特有的7个基因，与患者临床表现一致。 结论:本研究建立的9个基因多重PCR反应体系可快速准确的筛查Turner综合征患者是否携带Y染色体物质，具有检测成本低、操作简便、特异性强、检测周期短等优点，可供临床及时检出携带Y染色体物质的Turner综合征患者，为是否预防性切除性腺而防止恶性性腺肿瘤的发生提供诊断依据。

abstractsObjective:To develop a multiplex PCR method for a rapid detection of Y chromosome-specific sequences in patients with Turner syndrome.Methods:Nine genes were selected from various regions of the Y chromosome for designing the primers, which included SRY、TBL1Y、TSPY on the short arm of the Y chromosome, DDX3Y, HSFY1, RPS4Y2 and CDY1 on the long arm of Y chromosome and SHOX in the short arm and SPRY3 in the long arm of the pseudoautosomal region (PAR) of X and Y chromosomes. A multiplex PCR method for the nine genes in Y chromosome was established and optimized. The sensitivity was tested by using different amounts of genomic DNA. A total of 36 patients with Turner syndrome and a patient with male dwarfism with karyotype of 46, X, + mar were examined by the multiplex PCR method for the existence of materials from the Y chromosome. Results:The optimization results of the multiplex PCR reaction system (50 μL) showed that when the final concentration of upstream and downstream of each pair of primers was 0.1 μM, the multiplex PCR reaction of the 9 pairs of primers clearly amplified the target with the expected band size, and there was no non-specific amplification. The bands were clearly visible when the amount of genomic DNA in the multiple PCR reaction system was as low as 1 ng. By using the method, we have examined the 36 patients with Turner syndrome. One patient with Turner syndrome with karyotype of 45, X[40]/47XYY[21] amplified specific seven genes on Y chromosome, 35 patients with Turner syndrome amplified only two target genes SHOX and SPRY3, but not the other seven specific genes on the Y chromosome, which was in keeping with the clinical manifestations of such patients. Conclusion:This study established a multiplex PCR reaction system with nine genes, which can quickly and accurately screen Y chromosome materials in patients with Turner syndrome. It has the advantages of low cost, simple operation, high specificity and rapid turn-around time, and can be used to detect Turner syndrome patients with Y chromosome material in time. The method has provided a diagnostic basis for preventive gonad resection to prevent malignant gonadal tumors.