The Key Laboratory of Reproductive Genetics, Department of Obstetrics and Gynecology, Women's Hospital, School of Medicine, Zhejiang University, 1 Xueshi Road, Hangzhou, 310006, Zhejiang, China.
Department of Clinical Laboratory Research, Shulan Hospital, Zhejiang University, 848 Dongxin Road, Hangzhou, 310006, Zhejiang, China.
BMC Med Genomics. 2020 Jul 31;13(1):109. doi: 10.1186/s12920-020-00758-1.
The goal of this study was to determine whether Levey-Jennings charts, which are widely used in clinical laboratories, can be used to create standardized internal quality controls (IQCs) for prenatal molecular diagnosis.
Aneuploid amniocyte lines with trisomy 13, 21, and 18, and 47,XXY were established by transfection with SV40LTag-pcDNA3.1(-)and combined at different ratios to generate aneuploidy chimeric quality-control cell mixtures A to H. These quality-control cells were then used to calculate the [Formula: see text], [Formula: see text] ±1 standard deviation (SD), [Formula: see text] ±2 SD, and [Formula: see text] ±3 SD values to develop standardized IQCs for methods used for the prenatal diagnosis of aneuploidies such as FISH.
Methods for constructing aneuploid amniocyte lines were developed and a set of quality-control cells (A-H) were prepared. The [Formula: see text] ±1 SD, [Formula: see text] ±2 SD, and [Formula: see text] ±3 SD values of these quality-control cells for trisomy 13 and 21 were 10.2 ± 1.7, 10.2 ± 3.4, and 10.2 ± 5.1, and 90.3 ± 2.3, 90.3 ± 4.6, and 90.3 ± 6.9, respectively. Based on the values and Levey-Jennings charts, a set of standardized IQCs for prenatal diagnosis such as FISH were established.
This method resolves the problems of a shortage of quality-control materials and a lack of quality-control charts in prenatal molecular diagnosis such as NIPT, NGS, aCGH/SNP, PCR, and FISH. Levey-Jennings chart-based IQCs for prenatal diagnosis such as FISH can be used to easily monitor whether IQC results are within acceptable limits, and then infer whether the diagnostic results for clinical samples are reliable. We expect that this standardized IQC will be useful for a wide range of molecular diagnostic laboratories.
本研究旨在探讨临床实验室广泛应用的 Levey-Jennings 图表能否用于建立产前分子诊断的标准化内部质量控制（IQC）。
通过转染 SV40LTag-pcDNA3.1(-)，建立了三体 13、21 和 18、47，XXY 的不整倍体羊水细胞系，并以不同比例混合，生成不整倍体嵌合质控细胞混合物 A 至 H。然后使用这些质控细胞计算[Formula: see text]、[Formula: see text] ±1 标准差（SD）、[Formula: see text] ±2 SD 和 [Formula: see text] ±3 SD 值，为 FISH 等产前非整倍体诊断方法开发标准化 IQC。
建立了构建非整倍体羊水细胞系的方法，并制备了一组质控细胞（A-H）。这些质控细胞的三体 13 和 21 的[Formula: see text] ±1 SD、[Formula: see text] ±2 SD 和 [Formula: see text] ±3 SD 值分别为 10.2 ± 1.7、10.2 ± 3.4 和 10.2 ± 5.1，以及 90.3 ± 2.3、90.3 ± 4.6 和 90.3 ± 6.9。基于这些值和 Levey-Jennings 图表，建立了一套用于 FISH 等产前诊断的标准化 IQC。
该方法解决了 NIPT、NGS、aCGH/SNP、PCR 和 FISH 等产前分子诊断中质控材料短缺和质控图缺乏的问题。基于 Levey-Jennings 图表的 FISH 等产前诊断的 IQC 可用于轻松监测 IQC 结果是否在可接受范围内，进而推断临床样本的诊断结果是否可靠。我们预计这种标准化 IQC 将对广泛的分子诊断实验室有用。
