sequential testing sensitivity specificity question

SEQUENTIAL TESTING SENSITIVITY SPECIFICITY QUESTION DRIVERS

In contrast, studies using more challenging indications and early-stage samples have generated data that demonstrate significant room for improvement for the currently-available clinical assays in this setting 5, 7, 11. Publications have reported excellent agreement between cfDNA liquid biopsy testing and either tissue or blood-based PCR testing 8, 9, 10, primarily looking at NSCLC and colorectal cancer (CRC) in patients at advanced stage (Stage III-IV). The accuracy of liquid biopsy tests is an area of constant improvement and scientific debate. A better understanding of test performance and clinical utility and higher-accuracy methods are needed to broaden adoption of liquid biopsies as the standard of care in cancer treatment. Despite the successful commercial introduction of liquid biopsy testing in oncology 2, 3, 4, mutation detection performance limitations remain 5, 6, 7.

Current applications include replacing tissue testing when that is difficult or fails, re-testing as disease progresses on treatment, and earlier detection of drug resistance. Sequencing-based liquid biopsy testing offers tremendous promise for tailoring treatment regimens to the changing tumor genomic landscape. In previous work, we highlighted the complementarity between the RAS droplet digital polymerase chain reaction (ddPCR) method and NGS, which often confirmed ddPCR results and provided a larger overview of the major targetable alterations of 56 genes (56G oncology panel, Swift Biosciences, Ann Arbor, MI, USA) in one run at diagnosis with a 0.5% threshold in lung and colon cancers 1. Several techniques are currently available to detect cfDNA, including highly sensitive PCR (polymerase chain reaction) assays and next-generation sequencing (NGS). It is now possible to detect somatic alterations from minute cfDNA concentrations (less than 0.1%), enabling the detection of alterations in low-volume plasma samples. In fact, the use of cfDNA for sensitizing and resistance somatic mutation detection in oncodrivers for NSCLC was integrated into the European Medicines Agency (EMA) approval. Usually, these molecular mutational analyses are performed on formalin-fixed paraffin-embedded (FFPE) tissue samples at diagnosis or recurrence.īiopsies of advanced stage non-small cell lung cancer (NSCLC) need invasive exams in fragile patients, and therefore minimally invasive “liquid biopsies” have generated considerable enthusiasm. Molecular profiling of a cancer patient’s tumor to reveal targetable alterations is an important first step in the personalization of cancer treatment plans.

SEQUENTIAL TESTING SENSITIVITY SPECIFICITY QUESTION DRIVERS

Due to broader coverage and high sensitivity for insertions and deletions, sequencing via MAPs afforded important detection of additional actionable mutations.Ī growing understanding of cancer molecular complexity and the role of oncogenic drivers such as mutations in genes encoding the epidermal growth factor receptor ( EGFR), V-Ki-Ras2 Kirsten Rat Sarcoma 2 ( KRAS), (MET) and Anaplastic Lymphoma Kinase (ALK) genes have ushered in the era of targeted therapies. The accuracy of this cfDNA sequencing approach was similar to that of ddPCR assays in a clinical setting, down to an allele frequency of 0.1%. Non-actionable clonal hematopoiesis-associated variants were identified via sequencing in 21% of samples. MAPs sequencing detected somatic variants in 261 of 356 samples (73%). The ddPCR assay was used as the reference, since it is an established, accurate assay that can be performed contemporaneously on the same plasma sample. Sequencing of cfDNA via MAPs had a sensitivity of 98.5% and specificity 98.9%. This study reports data from 356 lung cancer patients receiving plasma testing as part of routine clinical management. We determined the accuracy of MAPs plasma sequencing with respect to droplet digital polymerase chain reaction assays (ddPCR), and tested whether actionable mutation discovery is improved by next-generation sequencing (NGS) in a clinical setting. Here, a new sequencing error-reduction method based on molecular amplification pools (MAPs) was utilized to analyze cfDNA in lung cancer patients. Circulating cell-free DNA (cfDNA) has the potential to be a specific biomarker for the therapeutic management of lung cancer patients.

Sequential testing sensitivity specificity question