Abstract 490: High-throughput mutational analysis in cell-free DNA by targeted next-generation sequencing

2016 ◽  
Author(s):  
Nga wan Rachel Tam ◽  
David Shames ◽  
Walter Darbonne
Keyword(s):  
Next Generation Sequencing ◽  
High Throughput ◽  
Mutational Analysis ◽  
Next Generation ◽  
Cell Free Dna ◽  
Targeted Next Generation Sequencing ◽  
Free Dna ◽  
Generation Sequencing

scholarly journals Somatic Tumor Mutations Detected by Targeted Next Generation Sequencing in Minute Amounts of Serum-Derived Cell-Free DNA

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Marjolein J. A. Weerts ◽  
Ronald van Marion ◽  
Jean C. A. Helmijr ◽  
Corine M. Beaufort ◽  
Niels M. G. Krol ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Next Generation ◽  
Cell Free Dna ◽  
Targeted Next Generation Sequencing ◽  
Free Dna ◽  
Generation Sequencing

scholarly journals Targeted next-generation sequencing of circulating cell-free DNA vs bone marrow in patients with acute myeloid leukemia

2020 ◽  
Vol 4 (8) ◽  
pp. 1670-1677
Author(s):  
Nicholas J. Short ◽  
Keyur P. Patel ◽  
Maher Albitar ◽  
Miguel Franquiz ◽  
Rajyalakshmi Luthra ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Acute Myeloid Leukemia ◽  
Next Generation Sequencing ◽  
Myeloid Leukemia ◽  
Next Generation ◽  
Cell Free Dna ◽  
Targeted Next Generation Sequencing ◽  
Free Dna ◽  
Generation Sequencing ◽  
Acute Myeloid

Abstract Circulating cell-free DNA (ccfDNA) allows for noninvasive peripheral blood sampling of cancer-associated mutations and has established clinical utility in several solid tumors. We performed targeted next-generation sequencing of ccfDNA and bone marrow at the time of diagnosis and after achieving remission in 22 patients with acute myeloid leukemia (AML). Among 28 genes sequenced by both platforms, a total of 39 unique somatic mutations were detected. Five mutations (13%) were detected only in ccfDNA, and 15 (38%) were detected only in bone marrow. Among the 19 mutations detected in both sources, the concordance of variant allelic frequency (VAF) assessment by both methods was high (R2 = 0.849). Mutations detected in only 1 source generally had lower VAF than those detected in both sources, suggesting that either method may miss small subclonal populations. In 3 patients, sequencing of ccfDNA detected new or persistent leukemia-associated mutations during remission that appeared to herald overt relapse. Overall, this study demonstrates that sequencing of ccfDNA in patients with AML can identify clinically relevant mutations not detected in the bone marrow and may play a role in the assessment of measurable residual disease. However, mutations were missed by both ccfDNA and bone marrow analyses, particularly when the VAF was <10%, suggesting that ccfDNA and bone marrow may be complementary in the assessment and monitoring of patients with AML.

Performance comparison of two next-generation sequencing panels to detect actionable mutations in cell-free DNA in cancer patients

2020 ◽  
Vol 58 (8) ◽  
pp. 1341-1348
Author(s):  
Mónica Macías ◽  
Eva Cañada-Higueras ◽  
Estibaliz Alegre ◽  
Arancha Bielsa ◽  
Javier Gracia ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Cancer Patients ◽  
Mutational Analysis ◽  
Performance Comparison ◽  
Next Generation ◽  
Advanced Cancer Patients ◽  
Cell Free Dna ◽  
Free Dna ◽  
Pan Cancer ◽  
Generation Sequencing

AbstractBackgroundGenomic alterations studies in cell-free DNA (cfDNA) have increasing clinical use in oncology. Next-generation sequencing (NGS) technology provides the most complete mutational analysis, but nowadays limited data are available related to the comparison of results reported by different platforms. Here we compare two NGS panels for cfDNA: Oncomine™ Pan-Cancer Cell-Free Assay (Thermo Fisher Scientific), suitable for clinical laboratories, and Guardant360® (GuardantHealth), with more genes targeted but only available in an outsourcing laboratory.MethodsPeripheral blood was obtained from 16 advanced cancer patients in which Guardant360® (G360) was requested as part of their clinical assistance. Blood samples were sent to be analyzed with G360 panel, and an additional blood sample was drawn to obtain and analyze cfDNA with Oncomine™ Pan-Cancer (OM) panel in an Ion GeneStudio S5™ System.ResultscfDNA analysis globally rendered 101 mutations. Regarding the 55/101 mutations claimed to be included by manufacturers in both panels, 17 mutations were reported only by G360, 10 only by OM and 28 by both. In those coincident cases, there was a high correlation between the variant allele fractions (VAFs) calculated with each panel (r = 0.979, p < 0.01). Regarding the six actionable mutations with an FDA-approved therapy reported by G360, one was missed with OM. Also, 12 mutations with clinical trials available were reported by G360 but not by OM.ConclusionsIn summary, G360 and OM can produce different mutational profile in the same sample, even in genes included in both panels, which is especially important if these mutations are potentially druggable.

scholarly journals 670. Rapid, Non-invasive Detection of Invasive Mycoplasma hominis Infection using the Karius Test, A Next-Generation Sequencing Test for Microbial Cell-free DNA in Plasma

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S390-S390
Author(s):  
Priya Edward ◽  
William V La Via ◽  
Mehreen Arshad ◽  
Kiran Gajurel
Keyword(s):  
Next Generation Sequencing ◽  
Case Series ◽  
Mycoplasma Hominis ◽  
Microbial Cell ◽  
Next Generation ◽  
Cell Free Dna ◽  
Non Invasive ◽  
Free Dna ◽  
Hominis Infection ◽  
Generation Sequencing

Abstract Background Mycoplasma hominis is typically associated with genital infections in women and is a rare cause of musculoskeletal infections often in immunocompromised hosts. Diagnosis of invasive Mycoplasma hominis infections are difficult due to challenges in culturing these organisms. Molecular diagnostics require an index of suspicion which may not be present at the time of tissue sampling. Accurate, rapid diagnosis of Mycoplasma hominis infections are important for antibiotic management. Methods Two cases of invasive Mycoplasma hominis infections are presented in which the Karius test (KT) was used to make the diagnosis. The KT is a CLIA certified/CAP-accredited next-generation sequencing (NGS) plasma test that detects microbial cell-free DNA (mcfDNA). After mcfDNA is extracted and NGS performed, human reads are removed and remaining sequences are aligned to a curated database of &gt; 1400 organisms. Organisms present above a statistical threshold are reported. Case review was performed for clinical correlation. Results A young woman with lupus nephritis status post renal transplant developed persistent fever with progressive multifocal culture-negative osteoarticular infection despite empiric ceftriaxone. An adolescent female presented with an ascending pelvic infection progressing to purulent polymicrobial peritonitis (see table) requiring surgical debridement and cefipime, metronidazole and micafungin therapy; her course was complicated by progressive peritonitis/abscesses. Karius testing detected high-levels of Mycoplasma hominis mcfDNA in both cases – at 3251 molecules/microliter (MPM) in the first case and 3914 MPM in the second case. The normal range of Mycoplasma hominis mcfDNA in a cohort of 684 normal adults is 0 MPM. The patients rapidly improved with atypical coverage with doxycycline and levofloxaxin. Clinical findings in 2 patients with M. hominis infection detected by the Karius Test Conclusion Open-ended, plasma-based NGS for mcfDNA provides a rapid, non-invasive method to diagnose invasive Mycoplasma hominis infection. This case series highlights the potential to diagnose infections caused by fastidious pathogens to better inform antimicrobial therapy and achieve favorable outcomes. Disclosures William V. La Via, MD, Karius (Employee)

scholarly journals Early solid tumor diagnosis through next-generation sequencing of cell-free DNA

2018 ◽  
Vol 12 (11) ◽  
pp. 1197-1201
Author(s):  
Demosthenes E Ziogas ◽  
Ioannis D Kyrochristos ◽  
Efstathios G Lykoudis ◽  
Dimitrios H Roukos
Keyword(s):  
Next Generation Sequencing ◽  
Solid Tumor ◽  
Tumor Diagnosis ◽  
Next Generation ◽  
Cell Free Dna ◽  
Free Dna ◽  
Generation Sequencing

scholarly journals Next-generation sequencing of microbial cell-free DNA for rapid noninvasive diagnosis of infectious diseases in immunocompromised hosts

F1000Research ◽  
2019 ◽  
Vol 8 ◽  
pp. 1194 ◽  
Author(s):  
Jose F. Camargo ◽  
Asim A. Ahmed ◽  
Martin S. Lindner ◽  
Michele I. Morris ◽  
Shweta Anjan ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Invasive Aspergillosis ◽  
Noninvasive Diagnosis ◽  
Cell Transplant ◽  
Next Generation ◽  
Cell Free Dna ◽  
Hematopoietic Stem ◽  
Free Dna ◽  
Immunocompromised Hosts ◽  
Generation Sequencing

Background: Cell-free DNA (cfDNA) sequencing has emerged as an effective laboratory method for rapid and noninvasive diagnosis in prenatal screening testing, organ transplant rejection screening, and oncology liquid biopsies but clinical experience for use of this technology in diagnostic evaluation of infections in immunocompromised hosts is limited.  Methods: We conducted an exploratory study using next-generation sequencing (NGS) for detection of microbial cfDNA in a cohort of ten immunocompromised patients with febrile neutropenia, pneumonia or intra-abdominal infection.  Results: Pathogen identification by cfDNA NGS demonstrated positive agreement with conventional diagnostic laboratory methods in 7 (70%) cases, including patients with proven/probable invasive aspergillosis, Pneumocystis jirovecii pneumonia, Stenotrophomonas maltophilia bacteremia, Cytomegalovirus and Adenovirus viremia. NGS results were discordant in 3 (30%) cases including two patients with culture negative sepsis who had undergone hematopoietic stem cell transplant in whom cfDNA testing identified the etiological agent of sepsis; and one kidney transplant recipient with invasive aspergillosis who had received >6 months of antifungal therapy prior to NGS testing. Conclusion: These observations support the clinical utility of measurement of microbial cfDNA sequencing from peripheral blood for rapid noninvasive diagnosis of infections in immunocompromised hosts. Larger studies are needed.

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