ColoGene

Performance Characteristics

• Entire test performed in-house

To establish analytical sensitivity and specificity, reference samples that contain both disease-associated and non-disease associated sequence variations were analyzed. Analytical sensitivity: The likelihood of the assay to detect a sequence variation when present within the analyzed genomic region (the test’s false negative rate) was established. All samples tested that have a positive result (“targeted mutation detected”) were correctly classified as positive after sequencing. Because the germline origin of the samples tested, the lower limit of detection was determined empirically as the allelic read percentage or allelic fraction (>35%, considering ideally an allelic read percentage of 50). Analytical specificity: The likelihood of the assay to detect only the target regions and not interfering regions, was established. The assay did not detect sequence variations when none were present within the analyzed genomic region (the test’s false positive rate). These parameters were established by comparing a platform test results to both methods that have been independently validated (Ion Torrent or Illumina, plus Sanger sequencing). Reportable range: The span of test result values over which the laboratory can establish the accuracy of the instrument, was established. There are areas of the targeted region that cannot be sequenced reliably and therefore are excluded from the reportable range. Unless otherwise indicated, all targeted regions were sequenced with >50x depth or supplemented with additional analysis. Reads were aligned to a reference sequence (GRCh37) and sequence changes were identified and interpreted in the context of a single clinically relevant transcript. Enrichment and analysis focus on the coding sequence of the indicated transcripts, 20 bp of flanking intronic sequence, and other specific genomic regions demonstrated to be causative of disease at the time of assay design. Promoters, untranslated regions and other non-coding regions, highly homologous regions and long homopolymers (>12nt) were not interrogated. All clinical significance observations were confirmed by orthogonal technologies. Reference range: GRCh37 was used as the reference genome against which the sequence reads were aligned and compared. It includes SNPs and other sequence variations that occur in the general population. For Illumina validation, annotations from the manufacturer’s software (Illumina-VariantStudio) was used for variant interpretation and population data analysis.

Is proficiency testing performed for this test? HelpWhether the test undergoes periodic internal or external evaluation of the accuracy of test results. Lab-provided.

No

Not provided