1. Cell Line Controls for the Genotyping of a Spectrum of Human Single Nucleotide Polymorphisms in the Clinical Laboratory
- Author
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Andrea Freystetter, Christian Paar, Joerg Berg, and Christine Kimbacher
- Subjects
Quality Control ,Genotype ,Genotyping Techniques ,Clinical Laboratory Techniques ,Reproducibility of Results ,HL-60 Cells ,Single-nucleotide polymorphism ,Clinical Laboratory Services ,Biology ,Polymorphism, Single Nucleotide ,Molecular biology ,General Biochemistry, Genetics and Molecular Biology ,Melting curve analysis ,Cell Line ,Polymorphism (computer science) ,Cell Line, Tumor ,Methylenetetrahydrofolate reductase ,biology.protein ,Humans ,SNP ,Allele ,K562 Cells ,Genotyping - Abstract
BACKGROUND Genotyping for clinically important single nucleotide polymorphisms (SNPs) is performed by many clinical routine laboratories. To support testing, quality controls and reference materials are needed. Those may be derived from residual patient samples, left over samples of external quality assurance schemes, plasmid DNA or DNA from cell lines. DNAs from cell lines are commutable and available in large amounts. METHODS DNA from 38 cell lines were examined for suitability as controls in 11 SNP assays that are frequently used in a clinical routine laboratory: FV (1691G>A), FII (20210G>A), PAI-1 4G/5G polymorphism, MTHFR (677C>T, 1298A>C), HFE (H63D, S65C, C282Y), APOE (E2, E3, E4), LPH (-13910C>T), UGT1A1 (*28, *36, *37), TPMT (*2, *3A, *3B, *3C), VKORC1 (-1639G>A, 1173C>T), CYP2C9 (*2, *3, *5). Genotyping was performed by real-time PCR with melting curve analysis and confirmed by bi-directional sequencing. RESULTS We find an almost complete spectrum of genotypic constellations within these 38 cell lines. About 12 cell lines appear sufficient as genotypic controls for the 11 SNP assays by covering almost all of the genotypes. However, hetero- and homozygous genotypes for FII and the alleles TPMT*2, UGT1A1*37 and CYP2C9*5 were not detected in any of the cell lines. CONCLUSIONS DNA from most of the examined cell lines appear suitable as quality controls for these SNP assays in the laboratory routine, as to the implementation of those assays or to prepare samples for quality assurance schemes. Our study may serve as a pilot to further characterize these cell lines to arrive at the status of reference materials.
- Published
- 2018
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