A novel use for Levey-Jennings charts in prenatal molecular diagnosis

Binghuan Weng,Lan Su,Jun Ying,Yanmei Yang ,Yali Xu ,Haokun Yang ,Min Chen

doi:10.1186/s12920-020-00758-1

Abstract

BackgroundThe 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.MethodsAneuploid 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 overline{mathrm{X}} , overline{mathrm{X}} ±1 standard deviation (SD), overline{mathrm{X}} ±2 SD, and overline{mathrm{X}} ±3 SD values to develop standardized IQCs for methods used for the prenatal diagnosis of aneuploidies such as FISH.ResultsMethods for constructing aneuploid amniocyte lines were developed and a set of quality-control cells (A-H) were prepared. The overline{mathrm{X}} ±1 SD, overline{mathrm{X}} ±2 SD, and overline{mathrm{X}} ±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.ConclusionsThis 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.

Highlights

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
With the widespread development of molecular diagnostic technologies such as aneuploidy detection by fluorescence in situ hybridization (FISH) [1], noninvasive prenatal testing (NIPT) [2], and QF-PCR technique [3], standardized quality control is urgently needed to ensure that prenatal molecular diagnoses are accurate and reliable [3,4,5]
We developed a set of aneuploidy chimeric quality-control cells (A-H) by immortalizing aneuploid amniocytes via SV40LTtransfection to address the lack of quality-control materials for methods used in prenatal diagnosis such as NIPT, next generation sequencing (NGS), and FISH [6,7,8]

Summary

Introduction

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. Over the past few decades, control charts have come to be widely used for IQC in clinical laboratories [10,11,12,13,14]. Levey and Jennings recommended that Shewhart’s control charts be used in clinical laboratories to provide a constant means of checking the reliability of the numerous tests run each day, and to make it possible to determine at a glance whether errors of analysis are beyond the permitted statistical variation defined by the procedure created in 1950 [10]. To date there have been few reports on the application of Levey-Jennings charts to prenatal molecular genetic diagnosis

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