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目的评估PCR法制备的人21/13α卫星探针进行荧光原位杂交(fluorescence in situ hybridization,FISH)的特异性及敏感性,并探讨和建立该探针的临床诊断价值和产前诊断方法。方法利用PCR方法制备的人21/13α卫星探针与中期淋巴细胞染色体杂交以确定其杂交位点特异性及敏感性;并将探针与50例妊娠16～26周抽取的未培养的羊水细胞进行FISH,同时进行常规细胞培养及染色体核型分析以检测该探针进行产前诊断的特异性和敏感性。结果探针与中期淋巴细胞染色体的FISH结果显示,杂交点位于21/13号染色体的着丝粒,正常组和唐氏组细胞21号和13号染色体的平均检出率为96%;探针与羊水细胞FISH结果显示,其检出率为85.6%,准确率100%。结论制备的人21/13α着丝粒特异DNA探针能简便、快速、准确的检测羊水细胞间期核中21号或13号染色体的数目的异常,该探针可用于Downs综合征与Patau综合征的诊断。
Objective To evaluate the specificity and sensitivity of fluorescence in situ hybridization (FISH) of human 21 / 13α satellite probe prepared by PCR and to explore and establish the clinical diagnostic value and prenatal diagnosis of this probe. Methods Hybridization of human 21 / 13α satellite probe with metaphase lymphocyte chromosome by PCR method was carried out to determine its hybridization site specificity and sensitivity. The probes were compared with 50 non-cultured amniotic fluid cells FISH was performed while routine cell culture and karyotype analysis were performed to examine the specificity and sensitivity of the probe for prenatal diagnosis. Results FISH results of probes and metaphase lymphocyte chromosomes showed that the hybridization sites were located on centromere of chromosome 21/13. The average detection rate of chromosome 21 and chromosome 13 was 96% in normal group and down-group cells. The probe FISH with amniotic fluid cells showed that the detection rate was 85.6%, the accuracy rate of 100%. Conclusions The human 21 / 13α centromere-specific DNA probe can be used to detect the abnormality of the number of chromosome 21 or 13 in interphalangeal amniotic fluid easily, rapidly and accurately. The probe can be used in the combination of Downs syndrome and Patau Diagnosis of the sign.