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首页> 外文期刊>Prenatal Diagnosis >A novel method for extracting DNA from chorionic villus samples for use in CVS-PCR, which ensures complete villus dissociation.
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A novel method for extracting DNA from chorionic villus samples for use in CVS-PCR, which ensures complete villus dissociation.

机译：一种从绒毛膜绒毛样品中提取DNA的新方法，用于CVS-PCR，可确保绒毛完全解离。

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OBJECTIVE: To demonstrate that glass disruption beads dissociate chorionic villus samples releasing DNA from mesenchymal and cytotrophoblast cells that is suitable for processing by CVS-PCR (rapid molecular aneuploidy testing). This method is quicker than conventional methods and may limit discrepancies between PCR and karyotype in certain types of placental mosaicism. METHOD: DNA was extracted from villus samples by mechanical disruption of the cells using glass beads. This method was compared to collagenase incubation followed by chelex extraction of the digested villus. PCR data generated were compared using standard criteria. RESULTS: DNA extracted by glass bead disruption generated data of equivalent quality to that obtained from DNA extracted using conventional collagenase and chelex-based extraction method. The case study demonstrates probable cytotrophoblast enrichment of a sample when processed by collagenase digestion and chelex incubation. Re-extraction of the digested sample by glass bead disruption resulted in cytotrophoblast and mesenchyme cells contributing to the supernatant. CONCLUSION: Glass bead disruption of chorionic villus samples is an effective, inexpensive and rapid DNA extraction method that dissociates villus ensuring that DNA from both cytotrophoblast and mesenchyme cells is represented in the supernatant. Extracted DNA produced is suitable for CVS-PCR and can be stored stably at - 20 degrees C. Copyright (c) 2008 John Wiley & Sons, Ltd.
机译：目的：证明玻璃破裂珠可分离绒毛膜绒毛样品，并从间充质和细胞滋养层细胞中释放DNA，适合通过CVS-PCR（快速分子非整倍性测试）进行处理。该方法比常规方法更快，并且可能会限制某些类型的胎盘镶嵌术中PCR和核型之间的差异。方法：使用玻璃珠通过机械破碎细胞从绒毛样品中提取DNA。将该方法与胶原酶温育相比较，然后酶切提取消化的绒毛。使用标准标准比较生成的PCR数据。结果：通过玻璃珠破裂提取的DNA产生的数据质量与使用常规胶原酶和基于chelex的提取方法提取的DNA相当。案例研究表明，通过胶原酶消化和Chelex孵育处理后，样品的细胞滋养层可能富集。通过玻璃珠破坏重新提取消化后的样品，导致滋养细胞和间充质细胞有助于上清液。结论：破坏绒毛膜绒毛样品的玻璃珠是一种有效，廉价且快速的DNA提取方法，该方法可分离绒毛，从而确保上皮细胞中存在来自滋养层细胞和间充质细胞的DNA。产生的提取DNA适用于CVS-PCR，可在20摄氏度下稳定保存。版权所有（c）2008 John Wiley＆Sons，Ltd.。

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