Garvin A M, Holzgreve W, Hahn S
Department of Obstetrics and Gynaecology and Laboratory for Prenatal Medicine, University of Basel,46 Schanzenstrasse, Basel CH-4031, Switzerland.
Nucleic Acids Res. 1998 Aug 1;26(15):3468-72. doi: 10.1093/nar/26.15.3468.
Single cell PCR is a powerful method for determining the genetic properties of individual cells. In the instance of heterozygous loci, however, preferential amplification of one allele can lead to allele drop out (ADO) of the other allele. Fortunately ADO does not occur in all amplifications, and is usually random when it does occur, with both alleles being equally susceptible to drop out. Therefore pooling of results from multiple independently amplified cells should greatly improve the analysis of diallelic loci, and the misdiagnosis rate of diallelic loci should decrease exponentially with the number of cells analysed. We have shown that this is true and that multiplex PCR allows for the simultaneous identification of a cell in a mixture of cells using microsatellite loci known to be informative, and accurate genotyping at other loci. This approach has practical applications to non-invasive prenatal diagnosis where small numbers of fetal cells in the presence of maternal cells must be both identified and genotyped at loci involved in genetic disease.
单细胞聚合酶链反应（PCR）是确定单个细胞遗传特性的一种强大方法。然而，在杂合基因座的情况下，一个等位基因的优先扩增可能导致另一个等位基因的等位基因缺失（ADO）。幸运的是，ADO并非在所有扩增中都会出现，并且当它发生时通常是随机的，两个等位基因对等位基因缺失的易感性相同。因此，汇集多个独立扩增细胞的结果应能大大改善双等位基因座的分析，并且双等位基因座的误诊率应随着分析细胞数量的增加而呈指数下降。我们已经证明情况确实如此，并且多重PCR允许使用已知具有信息性的微卫星基因座同时鉴定细胞混合物中的一个细胞，并在其他基因座进行准确的基因分型。这种方法在无创产前诊断中有实际应用，在无创产前诊断中，必须在涉及遗传疾病的基因座处识别并对存在于母体细胞中的少量胎儿细胞进行基因分型。
