A large-scale evaluation of amnio-PCR for the rapid prenatal diagnosis of fetal trisomy
A large-scale evaluation of amnio-PCR for the rapid prenatal diagnosis of fetal trisomy
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DOI：
10.1046/j.1469-0705.2001.00340.x
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发表时间：
2001-02-01
影响因子：
7.1
通讯作者：
Meredith, R
中科院分区：
文献类型：
--
作者：
Levett, LJ;Liddle, S;Meredith, R
目的传统的羊水染色体制备方法需要经过长时间的培养才能获得染色体。多重荧光定量聚合酶链反应（PCR）是一种无需细胞培养即可对DNA进行原位定量的分子生物学新技术。我们的目的是测试的可靠性，PCR使用胎儿DNA从羊水中的常见trisomies.Design的快速产前诊断（PCR），这是一个大的前瞻性研究5000例羊水穿刺标本。对21、18、13、X和Y染色体内的短串联重复序列进行特异性多重荧光定量PCR。所有羊膜穿刺标本随后进行分析，通过传统的核型分析methods.Results Amnio-PCR检测所有89个主要的常染色体三体在这个队列。性染色体异常的诊断是准确的情况下，涉及第一次减数分裂不分离。然而，进一步的标记是必要的，以检测性染色体异常所产生的第二次减数分裂不分离，突出使用特定的标记，使定量的X和Y染色体的重要性。结论快速产前诊断三体21，18，和13和性染色体异常，使用ANOPO-P CR是一种可靠的技术，有助于临床管理怀孕。该方法的速度将有助于最大限度地减少父母在等待诊断测试结果时的焦虑。
Objective Traditional chromosome preparation from amniotic fluid samples often involves lengthy culture procedures in order to obtain cells for analysis. Multiplex quantitative fluorescent polymerase chain reaction (PCR) is a new molecular biological technique capable of quantifying in-situ DNA without the need for cell culture. Our objective was to test the reliability of PCR using fetal DNA from amniotic fluid (amnio-PCR) for the rapid prenatal diagnosis of the common trisomies.Design This was a large prospective study of 5000 amniocentesis specimens. Multiplex quantitative fluorescent PCR was performed specifically for short tandem repeat sequences within chromosomes 21, 18, 13, X and Y. All aminocentesis samples were subsequently analyzed by traditional karyotyping methods.Results Amnio-PCR detected all 89 major autosomal trisomies in this cohort. Diagnosis of sex chromosome anomalies was accurate for cases involving first meiotic division nondisjunction. However, further markers were necessary to detect sex chromosome anomalies arising from second meiotic division nondisjunction, highlighting the importance of using specific markers that enable the quantification of both the X and the Y chromosomes simultaneously.Conclusions Rapid prenatal diagnosis of trisomies 21, 18, and 13 and the sex chromosome anomalies using amnio-P CR is a reliable technique that aids the clinical management of pregnancy. The speed of the methodology will help to minimize the period of parental anxiety in the wait for a diagnostic test result.