Preimplantation diagnosis of the beta1 integrin knockout mutation as a model for aneuploid gene testing.
Preimplantation diagnosis of the beta1 integrin knockout mutation as a model for aneuploid gene testing.
复制标题
β1 整合素敲除突变的植入前诊断作为非整倍体基因测试的模型。
DOI：
10.1007/s004390051134
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发表时间：
1999
期刊：
影响因子：
5.3
通讯作者：
Lebo,RV
中科院分区：
文献类型：
--
作者：
Kim,HS;Klimanskaya,IV;Damsky,CK;Pedersen,RA;Lebo,RV
选择常染色体β1整联蛋白基因敲除小鼠突变作为模型，以实验确定常染色体基因缺失和重复的着床前诊断试验的可靠性。在实验1中，分析了198个单独分解的单卵裂球，观察到的测试频率与从90%正常等位基因扩增、92%突变等位基因扩增、4%替代等位基因污染和4%未能将可扩增的靶DNA转移到PCR反应混合物中的独立导出值计算的数学预测频率相匹配。在144例表型正常的常染色体隐性遗传结果中，该实验正确预测了143例（99.3%）正常胚胎表型。实验2将单个活组织检查的卵裂球测试结果与培养1周直到滋养层生长的剩余胚胎细胞的测试结果进行比较。单活组织检查卵裂球分析正确预测了89个胚胎中的87个（98%）正常常染色体隐性表型，这些胚胎将被选择用于植入。实验3将两个单独检测的活组织检查的卵裂球的PCR结果与剩余培养的卵裂球的PCR结果进行比较，以提高检测的可靠性。考虑到胚胎只有在获得两个正常结果时才能植入，因此在44个测试的胚胎中，17个表型正常的胚胎中有17个将被植入。这些实验3的结果与数学预测一致，即植入两个未受影响的活组织检查卵裂球结果的胚胎中约99.9%将具有表型正常的基因型。
The autosomal β1 integrin knockout mouse mutation was selected as a model to experimentally determine preimplantation diagnosis test reliability for autosomal gene deletions and duplications. In experiment 1, which analyzed 198 individually disaggregated single blastomeres, the observed test frequencies matched the mathematically predicted frequencies calculated from the independently derived values of 90% normal allele amplification, 92% mutant allele amplification, 4% alternate allele contamination, and 4% failure to transfer amplifiable target DNA into the PCR reaction mix. This experiment correctly predicted a normal embryonic phenotype in 143 (99.3%) of the 144 phenotypically normal autosomal recessive results. Experiment 2 compared single biopsied blastomere test results to test results on the remaining embryonic cells cultured 1 week until trophoblast outgrowth. Single biopsied blastomere analysis correctly predicted a normal autosomal recessive phenotype in 87 (98%) of the 89 embryos that would have been selected for implantation. Experiment 3 compared the PCR results of two biopsied blastomeres tested independently to the PCR result from the remaining cultured blastomeres to improve test reliability. Given that embryos would have been implanted only when two normal results were obtained, 17 of 17 phenotypically normal embryos would have been implanted from among the 44 embryos tested. These experiment 3 results are consistent with the mathematical prediction that about 99.9% of embryos implanted with two unaffected biopsied blastomere results would have had a phenotypically normal genotype.