Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study
Next generation sequencing analysis is crucial for therapeutic decision in various solid tumor contexts. The sequencing method must remain accurate and robust throughout the instrument lifespan allowing the biological validation of patients’ results. This study aims to evaluate the long-term sequenc...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/13/5/937 |
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| author | Jessica Bamba-Funck Emmanuelle E. Fabre Marianne Kambouchner Olivier Schischmanoff |
| author_facet | Jessica Bamba-Funck Emmanuelle E. Fabre Marianne Kambouchner Olivier Schischmanoff |
| author_sort | Jessica Bamba-Funck |
| collection | DOAJ |
| description | Next generation sequencing analysis is crucial for therapeutic decision in various solid tumor contexts. The sequencing method must remain accurate and robust throughout the instrument lifespan allowing the biological validation of patients’ results. This study aims to evaluate the long-term sequencing performances of the Oncomine Focus assay kit allowing theranostic DNA and RNA variants detection on the Ion S5XL instrument. We evaluated the sequencing performances of 73 consecutive chips over a 21-month period and detailed the sequencing data obtained from both quality controls and clinical samples. The metrics describing sequencing quality remained stable throughout the study. We showed that an average of 11 × 10<sup>6</sup> (±0.3 × 10<sup>6</sup>) reads were obtained using a 520 chip leading to an average of 6.0 × 10<sup>5</sup> (±2.6 × 10<sup>5</sup>) mapped reads per sample. Of 400 consecutive samples, 95.8 ± 16% of amplicons reached the depth threshold of 500X. Slight modifications of the bioinformatics workflow improved DNA analytical sensitivity and allowed the systematic detection of expected SNV, indel, CNV, and RNA alterations in quality controls samples. The low inter-run variability of DNA and RNA—even at low variant allelic fraction, amplification factor, or reads counts—indicated that our method was adapted to clinical practice. The analysis of 429 clinical DNA samples indicated that the modified bioinformatics workflow allowed detection of 353 DNA variants and 88 gene amplifications. RNA analysis of 55 clinical samples revealed 7 alterations. This is the first study showing the long-term robustness of the Oncomine Focus assay in clinical routine practice. |
| first_indexed | 2024-03-11T07:27:21Z |
| format | Article |
| id | doaj.art-1ad3395314d14de4bc9379682fb7b219 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-11T07:27:21Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-1ad3395314d14de4bc9379682fb7b2192023-11-17T07:30:12ZengMDPI AGDiagnostics2075-44182023-03-0113593710.3390/diagnostics13050937Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life StudyJessica Bamba-Funck0Emmanuelle E. Fabre1Marianne Kambouchner2Olivier Schischmanoff3Laboratory of Biochemistry, Hôpital Avicenne, Hôpitaux Universitaires Paris-Seine-Saint-Denis, Assistance Publique—Hôpitaux de Paris, F-93000 Bobigny, FranceLaboratory of Biochemistry, Hôpital Avicenne, Hôpitaux Universitaires Paris-Seine-Saint-Denis, Assistance Publique—Hôpitaux de Paris, F-93000 Bobigny, FranceDepartment of Pathology, Hôpital Avicenne, Hôpitaux Universitaires Paris-Seine-Saint-Denis, Assistance Publique—Hôpitaux de Paris, F-93000 Bobigny, FranceLaboratory of Biochemistry, Hôpital Avicenne, Hôpitaux Universitaires Paris-Seine-Saint-Denis, Assistance Publique—Hôpitaux de Paris, F-93000 Bobigny, FranceNext generation sequencing analysis is crucial for therapeutic decision in various solid tumor contexts. The sequencing method must remain accurate and robust throughout the instrument lifespan allowing the biological validation of patients’ results. This study aims to evaluate the long-term sequencing performances of the Oncomine Focus assay kit allowing theranostic DNA and RNA variants detection on the Ion S5XL instrument. We evaluated the sequencing performances of 73 consecutive chips over a 21-month period and detailed the sequencing data obtained from both quality controls and clinical samples. The metrics describing sequencing quality remained stable throughout the study. We showed that an average of 11 × 10<sup>6</sup> (±0.3 × 10<sup>6</sup>) reads were obtained using a 520 chip leading to an average of 6.0 × 10<sup>5</sup> (±2.6 × 10<sup>5</sup>) mapped reads per sample. Of 400 consecutive samples, 95.8 ± 16% of amplicons reached the depth threshold of 500X. Slight modifications of the bioinformatics workflow improved DNA analytical sensitivity and allowed the systematic detection of expected SNV, indel, CNV, and RNA alterations in quality controls samples. The low inter-run variability of DNA and RNA—even at low variant allelic fraction, amplification factor, or reads counts—indicated that our method was adapted to clinical practice. The analysis of 429 clinical DNA samples indicated that the modified bioinformatics workflow allowed detection of 353 DNA variants and 88 gene amplifications. RNA analysis of 55 clinical samples revealed 7 alterations. This is the first study showing the long-term robustness of the Oncomine Focus assay in clinical routine practice.https://www.mdpi.com/2075-4418/13/5/937next generation sequencing (NGS)solid tumorssingle nucleotide variation (SNV)copy number variation (CNV)fusion transcriptlong-term follow-up |
| spellingShingle | Jessica Bamba-Funck Emmanuelle E. Fabre Marianne Kambouchner Olivier Schischmanoff Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study Diagnostics next generation sequencing (NGS) solid tumors single nucleotide variation (SNV) copy number variation (CNV) fusion transcript long-term follow-up |
| title | Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study |
| title_full | Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study |
| title_fullStr | Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study |
| title_full_unstemmed | Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study |
| title_short | Performance Characteristics of Oncomine Focus Assay for Theranostic Analysis of Solid Tumors, A (21-Months) Real-Life Study |
| title_sort | performance characteristics of oncomine focus assay for theranostic analysis of solid tumors a 21 months real life study |
| topic | next generation sequencing (NGS) solid tumors single nucleotide variation (SNV) copy number variation (CNV) fusion transcript long-term follow-up |
| url | https://www.mdpi.com/2075-4418/13/5/937 |
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