دورية أكاديمية
A Novel System Control for Quality Control of Diagnostic Tests Based on Next-Generation Sequencing.
| العنوان: | A Novel System Control for Quality Control of Diagnostic Tests Based on Next-Generation Sequencing. |
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| المؤلفون: | Chan M; Vela Research, Singapore., Smirnov A; Vela Research, Singapore., Mulawadi F; Vela Research, Singapore., Lim P; Vela Research, Singapore., Lim WH; Vela Research, Singapore., Leong ST; Vela Research, Singapore., Low HM; Vela Research, Singapore., Yee MQ; Vela Research, Singapore., Yeo YQ; Vela Research, Singapore., Zhou X; Vela Research, Singapore., Lee C; Vela Research, Singapore., Huang W; Vela Research, Singapore., Welebob L; Vela Research, Singapore., Wu M |
| المصدر: | The journal of applied laboratory medicine [J Appl Lab Med] 2016 Jul 01; Vol. 1 (1), pp. 25-35. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Oxford University Press Country of Publication: England NLM ID: 101693884 Publication Model: Print Cited Medium: Print ISSN: 2576-9456 (Print) Linking ISSN: 24757241 NLM ISO Abbreviation: J Appl Lab Med Subsets: PubMed not MEDLINE; MEDLINE |
| أسماء مطبوعة: | Publication: 2020- : Oxford : Oxford University Press Original Publication: [Washington, DC] : American Association for Clinical Chemistry, [2016]-[2019] |
| مستخلص: | Background: We describe a novel system control (SC) implemented in an automated AmpliSeq™-based next-generation sequencing (NGS)2 run that simultaneously acts as (a) an external positive/sensitivity control, (b) a spike-in QC for DNA extraction, and (c) a nontemplate control to detect exogenous DNA contamination. Methods: Plasmids carrying wild-type tobacco mosaic virus sequence and a sequence with three designed mutations were synthesized and mixed, such that the mutations are present at 5% variant frequency in the mixture designated as SC. SC was used as a stand-alone sample and spiked into each sample in each run. A cell line-derived reference material, in both a formalin-fixed paraffin-embedded (FFPE) sample and genomic DNA (gDNA), was sequenced in the same runs. Results: By interpolation, 100 fg SC spiked in FFPE sample produced sequencing coverage equivalent to approximately 3 fg in the gDNA. In the SC-only sample, all three designed mutations were recovered around 5% as expected, while no significant reads of human genome were present. In samples with a common PCR inhibitor, coverage for both SC and target amplicons were eliminated. An inverse relationship between the coverage of SC and DNA input was observed. In clinical samples, the ratio of SC to the median coverage of sample can be used to indicate insufficient DNA input. Conclusions: The SC is an elegant and comprehensive QC concept for NGS-based diagnostic tests. (© 2016 by American Association for Clinical Chemistry.) |
| تواريخ الأحداث: | Date Created: 20210225 Latest Revision: 20210225 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1373/jalm.2016.020131 |
| PMID: | 33626812 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2576-9456 |
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| DOI: | 10.1373/jalm.2016.020131 |