PR7000-1421 HP | Alldatasheet
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Technical content
A simple and robust CE-Marked diagnostic assay for complete mutation analysis of the CFTR gene by Next-Generation Sequencing. Mutations of the CFTR gene affect functioning of the chloride ion channels in epithelial cell membranes, leading to cystic fibrosis and other CFTR-related diseases. 100% 90% 0 0,05 0,1 0,15 0,2 0,25 0,3 0,35 0,4 0,45 0,5 80% 70% 60% 50% 40% 30% 20% 10% Percentage of amplicons x mean coverage Graph representing the read counts for all 48 CFTR MASTR Dx amplicons, showing their uniform representation. To allow comparison between samples, the read counts were normalized. validated for dried blood spots Application
- For first or second line testing in your CFTR work-up
- For whole blood & dried blood spots derived DNA
- For complete mutational spectrum analysis Gene analyzed CFTR Genomic region analyzed 8.7 kb All exons ± 30 bp flanking region, selected introns, part of promoter Number of amplicons 48 including 11 control amplicons Amplicon length 300-450 bp Number of plexes 2 DNA amount required 20 ng per multiplex reaction Validated (CE-IVD) with MiSeq® System, Illumina Mutations included All exonic SNVs & small indels CNVs* ~30 bp of intronic flanking sequence HP variants intron 9 (8): c.1210-34TG(m); c.1210-12T(n) Selected deep intronic regions: intron 7 (6b): c.1002-1110_1113delTAAG c.1002-1111A>C intron 12 (11): c.1679+1634A>G (1811+1.6kbA->G) c.1680-883A>G intron 22 (19): c.3718-2477C>T (3849+10kbC->T Promoter region up to position c.-108 DNA extracted from blood DNA extracted from dried blood spots On target read pair counts 98.92 % [92.70 %-99.51 %] 99.90 % [99.53 %-99.98 %] Uniformity of amplification (0.2x mean coverage) 97.83 % 100 % *For Research Use Only. Not for use in diagnostic procedures CFTR MASTR Dx on MiSeq, germline DNA
Cat. No. Product Name Reactions PR Number MR-2021.024 CFTR MASTR Dx PR7000-1421 MR-2021.048 48 MID (Molecular Identifiers) kits are necessary to complete the workflow. For In Vitro Diagnostic Use. *For Research Use Only. Not for use in diagnostic procedures Extracted from blood Extracted from dried blood spots Performance parameter Observed [95 % CI] Using JSI SeqNext Observed [95 % CI] Using Sophia DDM Observed Using JSI SeqNext Observed Using Sophia DDM Sensitivity 100 % [ ≥ 99.217 %] 100 % Specificity 99.990 % 100 % [ ≥ 99.999 %] 1438/1438 = 100 % 1438/1438 = 100 % Accuracy 99.990% 100 % [ ≥ 99.999 %] Repeatability & reproducibility 99.997 % 100 % [ ≥ 99.990 %] Supporting publications: Applicability and Efficiency of NGS in Routine Diagnosis: In-Depth Performance Analysis of a Complete Workflow for CFTR Mutation Analysis. Andrien Pagin et al. (2016) PLOS ONE DOI: 10.1371/journal.pone.0149426, 22 Feb 2016* *This study was done by a third party and was verified by Multiplicom. Advised maximum number of samples per run CFTR MASTR Dx on MiSeq; germline DNA WorkflowPublications Performance Characteristics Sequencing System Illumina MiSeq® Reagent Kit Flow cell Nano v 2 2 x 251 cycles 2 x 251 cycles 2 x 276 cycles SNV variant calling Minimal coverage per allele: 50 52 786 1441 SNV and CNV* variant calling Minimal coverage per amplicon: 200 26 393 720 Plan run Multiplex PCR Quality Control Universal PCR Quality Control Equimolar poolingMixing plexes of single sample Purifi cation Multiplex PCR Dilution Universal PCR Mixing Clean up Pooling SEQUENCING DATA ANALYSIS MASTR workfl ow NGS 5991-8404EN Printed in Belgium, September 2017 1709 - EU