论文部分内容阅读
目的探讨SYBRGreenⅠ实时荧光PCR技术用于非整倍体诊断的可行性。方法选择13号染色体上的ABCC4基因、18号染色体上的TYMS基因、21号染色体上的DSCR3基因、X染色体上的HPRT2基因和Y染色体上的SRY基因为目的基因,以12号染色体上的管家基因GAPDH为参照基因,选取SYBRGreenⅠ作为荧光染料,采用双标准曲线相对定量PCR法进行检测,并与核型分析结果进行比对。结果检测13号染色体时,计算目的基因与参照基因比值,正常标本组(0.90±0.31)与三体标本组(1.39±0.12)的差别有显著意义(P=0.003);检测18号染色体时,正常标本组(1.07±0.44)与三体标本组(1.66±0.12)的差别有显著意义(P=0.000);检测21号染色体时,正常标本组(0.84±0.27)与三体标本组(1.73±0.54)的差别有显著意义(P=0.000);检测X染色体时,45,X组(0.62±0.12)与46,XY组(0.63±0.25)比值无差别(P=0.965),46,XX组(1.32±0.37)与47,XXY组(1.20±0.35)比值无差别(P=0.326),单个拷贝X(包括45,X、46,XY)(0.63±0.23)与两个拷贝X(46,XX、47,XXY)(1.26±0.36)比值差别有统计学意义(P=0.000);检测Y染色体时,正常女性(46,XX)无目的基因扩增,正常男性组(46,XY)(1.57±0.54)与三体标本组(47,XYY)(3.08±0.15)比值差别有统计学意义(P=0.003)。结论SYBRGreenⅠ实时荧光定量技术可用于快速诊断染色体非整倍体。
Objective To investigate the feasibility of SYBRGreen Ⅰ real-time fluorescence PCR for aneuploidy diagnosis. Methods The ABCC4 gene on chromosome 13, the TYMS gene on chromosome 18, the DSCR3 gene on chromosome 21, the HPRT2 gene on chromosome X and the SRY gene on chromosome Y were selected as the target genes. Gene GAPDH as a reference gene, select SYBRGreen I as a fluorescent dye, using double-standard curve relative quantitative PCR method for detection and karyotype analysis results were compared. Results When the chromosome 13 was detected, the ratio of the target gene to the reference gene was calculated. The difference between the normal group (0.90 ± 0.31) and the trisomy group (1.39 ± 0.12) was significant (P = 0.003) The difference between normal group (1.07 ± 0.44) and trisomic group (1.66 ± 0.12) was significant (P = 0.000). On chromosome 21, the normal group (0.84 ± 0.27) and trisomic group ± 0.54) (P = 0.000). There was no significant difference (P = 0.965), 46, XX when the X chromosome was detected in the group of 45, X (0.62 ± 0.12) and 46, XY There was no significant difference between the two groups (1.32 ± 0.37) and 47, XXY group (1.20 ± 0.35) (P = 0.326) (46, XX) in the normal male group (46, XY), the normal male group (46, XY) (1.57 ± 0.54) and trisomy (47, XYY) (3.08 ± 0.15), there was a significant difference (P = 0.003). Conclusion SYBRGreen Ⅰ real-time fluorescence quantitative assay can be used to rapidly diagnose aneuploidy in chromosomes.