前瞻性验证定量荧光聚合酶链反应快速检测常见非整倍体。

Prospective validation of quantitative fluorescent polymerase chain reaction for rapid detection of common aneuploidies.

机构信息

Genetics Program, North York General Hospital, 4001 Leslie Street, Toronto, Ontario, Canada.

出版信息

Genet Med. 2011 Feb;13(2):140-7. doi: 10.1097/GIM.0b013e3182036763.

DOI:10.1097/GIM.0b013e3182036763

PMID:21233717

Abstract

PURPOSE

To prospectively validate a quantitative fluorescent polymerase chain reaction (PCR) assay as a method of rapid prenatal aneuploidy detection for chromosomes 13, 18, 21, X, and Y.

METHODS

A commercial quantitative fluorescent PCR kit was validated on 200 known, blinded, prenatal DNA specimens. The kit was then validated prospectively on 1069 amniotic fluid specimens, and the results were compared with the karyotype results and the results of interphase fluorescence in situ hybridization testing, when performed in the course of standard care. Turnaround time was monitored in a subset of the prospective specimens.

RESULTS

The analytical sensitivity and specificity of testing in the validation specimens were 98.9% and 100%, respectively. There were no false positives and a single false negative, a mosaic sex chromosome aneuploidy interpreted as normal. In the prospective study, the analytical sensitivity and specificity were 98% and 100%, respectively. No false positives and a single false negative, again a sex chromosome mosaic, were detected. Overall, 72.5% of all chromosomal anomalies and 87.7% of clinically significant chromosome anomalies were detected by quantitative fluorescent PCR. The average and median turnaround times were 30.5 and 25.1 hours, respectively.

CONCLUSIONS

Quantitative fluorescent PCR is a robust and accurate method of rapid prenatal aneuploidy detection.

摘要

目的

前瞻性验证一种定量荧光聚合酶链反应（PCR）检测方法，作为快速产前非整倍体检测 13、18、21、X 和 Y 染色体的方法。

方法

在 200 份已知的、盲法的产前 DNA 标本上验证了一种商业定量荧光 PCR 试剂盒。然后，该试剂盒在 1069 份羊水标本中进行了前瞻性验证，并将结果与核型结果和间期荧光原位杂交检测结果进行了比较，这些结果是在标准护理过程中进行的。在部分前瞻性标本中监测了周转时间。

结果

在验证标本中的检测分析灵敏度和特异性分别为 98.9%和 100%。没有假阳性和一个假阴性，一个解释为正常的镶嵌性性染色体非整倍体。在前瞻性研究中，分析灵敏度和特异性分别为 98%和 100%。没有假阳性和一个假阴性，又是一个性染色体镶嵌体。总的来说，定量荧光 PCR 检测到了 72.5%的所有染色体异常和 87.7%的临床显著染色体异常。平均和中位数的周转时间分别为 30.5 和 25.1 小时。