Hessner M J, Agostini T A, Bellissimo D B, Endean D J, Pircon R A, Kirschbaum N E
Clinical Laboratories, Blood Center of Southeastern Wisconsin, Milwaukee 53201, USA.
Am J Obstet Gynecol. 1997 Feb;176(2):327-33. doi: 10.1016/s0002-9378(97)70493-9.
Fetuses at risk for immune cytopenic disorders can be identified by molecular genotyping assays. To better understand the impact of maternal contamination on genotyping results, the levels of contamination that are routinely encountered during prenatal testing of fetal samples and the sensitivity of allele-specific polymerase chain reaction in detecting paternal alloalleles were examined.
Reconstitution experiments were performed to define the sensitivity of allele-specific polymerase chain reaction assays. The sensitivities of allele-specific polymerase chain reactions and polymerase chain reaction-restriction fragment length polymorphism were compared for detection of the factor V Leiden mutation.
A quantitative analysis of variable-number tandem repeat loci revealed maternal contamination in 4 of 56 fetal samples. Contaminating deoxyribonucleic acid compromised genotyping results when it comprised between 94% and 99% of the total deoxyribonucleic acid. Allele-specific polymerase chain reaction was found to be the more sensitive technique (0.8% sensitivity vs 13% sensitivity).
These results illustrate that allele-specific polymerase chain reaction is well suited for reliable prenatal identification of fetuses at risk of immune cytopenic disorders.
可通过分子基因分型检测来识别有免疫性血细胞减少症风险的胎儿。为了更好地了解母体污染对基因分型结果的影响，对胎儿样本产前检测中常规遇到的污染水平以及等位基因特异性聚合酶链反应检测父本等位基因的灵敏度进行了研究。
进行了重组实验以确定等位基因特异性聚合酶链反应检测的灵敏度。比较了等位基因特异性聚合酶链反应和聚合酶链反应-限制性片段长度多态性检测因子V Leiden突变的灵敏度。
对可变数目串联重复序列位点的定量分析显示，56份胎儿样本中有4份存在母体污染。当污染的脱氧核糖核酸占总脱氧核糖核酸的94%至99%时，会影响基因分型结果。发现等位基因特异性聚合酶链反应是更灵敏的技术（灵敏度为0.8%，而另一技术为13%）。
这些结果表明，等位基因特异性聚合酶链反应非常适合可靠地产前识别有免疫性血细胞减少症风险的胎儿。
