Here we compared clonotype identification by allele-specific oligonucleotide real-time quantitative-PCR (ASO RQ-PCR) and next-generation sequencing (NGS) in 80 multiple myeloma patients. ASO RQ-PCR was applicable in 49/55 (89%) and NGS in 62/78 (80%). Clonotypes identified by both methods were identical in 33/35 (94%). Sensitivity of 10 was confirmed in 28/29 (96%) by NGS while sensitivity of RQ-PCR was 10 in 7 (24%), 5 × 10 in 15 (52%), and 10 in 7 (24%). Among 14 samples quantifiable by ASO RQ-PCR, NGS yielded comparable results in 12 (86%). Applicability of NGS can be improved if immunoglobulin heavy-chain incomplete DJ primers are included.
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http://dx.doi.org/10.3389/fonc.2020.611021 | DOI Listing |
Front Oncol
January 2021
Department of Medicine, Queen Mary Hospital, The University of Hong Kong, Pokfulam, Hong Kong.
Here we compared clonotype identification by allele-specific oligonucleotide real-time quantitative-PCR (ASO RQ-PCR) and next-generation sequencing (NGS) in 80 multiple myeloma patients. ASO RQ-PCR was applicable in 49/55 (89%) and NGS in 62/78 (80%). Clonotypes identified by both methods were identical in 33/35 (94%).
View Article and Find Full Text PDFJ Clin Exp Hematop
December 2020
Niigata University of Pharmacy and Applied Life Sciences, Faculty of Pharmaceutical Sciences, Department of Pathophysiology, Niigata, Japan.
Treatment outcomes of chronic lymphocytic leukemia (CLL) have improved since chemoimmunotherapy and novel drugs became available for CLL treatment; therefore, more sensitive methods to evaluate residual CLL cells in patients are required. Measurable residual disease (MRD) has been assessed in several clinical trials on CLL using flow cytometry, real-time quantitative PCR (RQ-PCR) with allele-specific oligonucleotide (ASO) primers, and high-throughput sequencing. MRD assessment is useful to predict the treatment outcomes in the context of chemotherapy and treatment with novel drugs such as venetoclax.
View Article and Find Full Text PDFFront Oncol
August 2019
Hematology, Department of Translational and Precision Medicine, Sapienza University of Rome, Rome, Italy.
In chronic lymphocytic leukemia (CLL), there is a growing interest for minimal residual disease (MRD) monitoring, due to the availability of drug combinations capable of unprecedented complete clinical responses. The standardized and most commonly applied methods to assess MRD in CLL are based on flow cytometry (FCM) and, to a lesser extent, real-time quantitative PCR (RQ-PCR) with allele-specific oligonucleotide (ASO) primers of immunoglobulin heavy chain genes (IgH). Promising results are being obtained using droplet digital PCR (ddPCR) and next generation sequencing (NGS)-based approaches, with some advantages and a potential higher sensitivity compared to the standardized methodologies.
View Article and Find Full Text PDFFront Oncol
August 2019
Istituto di Ematologia "L.A.Seràgnoli," Azienda Ospedaliera Sant'Orsola-Malpighi, Bologna, Italy.
There is an increasing clinical interest in the measure and achievement of minimal residual disease (MRD) negativity in the bone marrow of Multiple Myeloma (MM) patients, as defined equally either by Multicolor Flow Cytometry (MFC) or by Next Generation Sequencing (NGS) technologies. At present, modern technologies allow to detect up to one on 104 or on 105 or even on 106 cells, depending on their throughput. MFC approaches, which have been progressively improved up to the so-called Next Generation Flow (NGF), and NGS, which proved clear advantages over ASO-PCR, can detect very low levels of residual disease in the BM.
View Article and Find Full Text PDFHematology
December 2019
b Department of Medicine , The University of Hong Kong, Hong Kong , People's Republic of China.
Objective: Buffy coat and ficoll of bone marrow (BM) are viable options for the study of minimal residual disease (MRD) in multiple myeloma (MM). As yet, there is no data about the superiority of either sample types. Herein, we aimed to address this issue.
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