Development and implementation of a new rapid aneuploidy diagnostic service within the UK National Health Service and implications for the future of prenatal diagnosis
Development and implementation of a new rapid aneuploidy diagnostic service within the UK National Health Service and implications for the future of prenatal diagnosis
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DOI：
10.1016/s0140-6736(01)06183-9
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发表时间：
2001-09-29
期刊：
影响因子：
168.9
通讯作者：
Ogilvie, CM
中科院分区：
文献类型：
--
作者：
Mann, K;Fox, SP;Ogilvie, CM
背景 染色体异常的产前诊断通常通过对培养细胞的染色体进行全面核型分析来进行。孕妇平均需要等待 13-14 天才能得到结果。常染色体三体性约占显着异常的 80%，可通过定量荧光 (QF) PCR 进行检测。我们报告了这项技术的开发和实施，作为英国国家卫生服务 (NHS) 诊断部门内第一个此类常规服务。方法我们设计了一种“单管测试”，包括 21 号染色体上的多态性四核苷酸重复序列的 4 个引物对、18 号染色体上的序列的 4 个引物对、13 号染色体上的序列的 3 个引物对和一个用于识别性染色体的引物对。我们的 NHS 诊断部门在 2000 年 4 月至 2001 年 4 月期间收到的所有产前样本均经过检测。 DNA提取后，进行PCR扩增，并在基于毛细管的遗传分析仪上分离产物；结果用专用软件解释。对所有样本进行后续核型分析。结果检测了1148份羊水样本、188份绒毛膜绒毛样本和37份胎儿组织样本；使用我们当前的方案，扩增失败率为零。获得并报告了 1314 份 (98%) 产前样本的 QF-PCR 结果；其余 22 例 (2%) 由于母体细胞污染而无法提供信息。在绒毛膜绒毛样本中发现了一例镶嵌现象，以及两例表明四核苷酸重复体细胞扩张的情况。没有获得假阳性或假阴性结果。最后 4 个月数据收集的平均报告时间为 1.25 个工作日。 解释 QF-PCR 非整倍体检测是一种高效、准确的技术，用于检测产前样本中的常染色体三体性。这项服务的实施可以快速诊断异常情况，并尽早让女性放心并获得正常结果。
Background Prenatal diagnosis for chromosome abnormality is routinely undertaken by full karyotype analysis of chromosomes from cultured cells; pregnant women must wait on average 13-14 days for their results. Autosomal trisomies, which account for around 80% of significant abnormalities, can be detected by quantitative fluorescence (QF) PCR. We report on the development and implementation of this technique as the first such routine service within a diagnostic department of the UK National Health Service (NHS).Methods We designed a "one-tube test" comprising four primer pairs for polymorphic tetranucleotide repeat sequences on chromosome 21, four primer pairs for sequences on chromosome 18, three primer pairs for sequences on chromosome 13, and one primer pair to identify the sex chromosomes. All prenatal samples received by our NHS diagnostic department between April, 2000, and April, 2001, were tested. After DNA extraction, PCR amplification was done and the products separated on a capillary-based genetic analyser; the results were interpreted with dedicated software. Follow-up karyotype analysis was done on all samples.Findings 1148 amniotic fluid samples, 188 chorionic villus samples, and 37 fetal tissue samples were tested; the amplification failure rate was zero with our current protocol. QF-PCR results were obtained and reported on 1314 (98%) of the prenatal samples; the remaining 22 (2%) were uninformative because of maternal-cell contamination. One case of mosaicism in a chorionic villus sample, and two cases indicating somatic expansion of a tetranucleotide repeat were found. No false positive or false negative results were obtained. The mean reporting time for the last 4 months of data collection was 1.25 working days.Interpretation QF-PCR aneuploidy testing is an efficient and accurate technique for the detection of autosomal trisomies in prenatal samples. Implementation of this service has led to the rapid diagnosis of abnormalities and early reassurance for women with normal results.