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Detection of maternal cell contamination in amniotic fluid cell cultures using fluorescent labelled microsatellites.

机译:使用荧光标记的微卫星检测羊水细胞培养物中的母体细胞污染。

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摘要

A rapid PCR based assay was used to ascertain the presence of maternal cell contamination (MCC) in amniotic fluid cell cultures and to exclude MCC in cases where cytogenetic analysis was possible only from one primary cell culture. Six 6-carboxyfluorescein (FAM) and three 6-carboxyfluorescein hexachloride (HEX) labelled primer sets were used to amplify two tetra- and seven dinucleotide repeat polymorphisms. The PCR amplifications were multiplexed in (three) three primer set reactions and visualised on an Applied Biosystems 373A sequencer running Genescan 672 software. The microsatellite products obtained from 200 amniotic fluid cell cultures where the karyotype was female were compared against corresponding maternal blood PCR products. A single case of MCC was detected indicating the usefulness of such assays. We suggest that screening for MCC should be considered in instances where the amniotic fluid sample is bloodstained or was obtained with difficulty, or where the karyotype is female and chromosome analysis is not possible from more than one primary cell culture.
机译:基于快速PCR的测定可用于确定羊水细胞培养物中母体细胞污染(MCC)的存在,在仅可从一种原代细胞培养物中进行细胞遗传学分析的情况下,可排除MCC。六个6-羧基荧光素(FAM)和三个6-羧基荧光素六氯化物(HEX)标记的引物组用于扩增两个四核苷酸和七个二核苷酸重复多态性。将PCR扩增物在(三个)三个引物组反应中多路复用,并在运行Genescan 672软件的Applied Biosystems 373A测序仪上可视化。比较了从200个羊水细胞培养物中获得的微卫星产物,其中核型为雌性,与相应的母体血液PCR产物进行了比较。检测到一例MCC,表明此类测定法的有用性。我们建议在羊水样本被染血或难以获得羊膜样本的情况下,或核型为女性且无法从一种以上的原代细胞培养物中进行染色体分析的情况下,应考虑筛查MCC。

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