High Sensitivity Quantitative Allele Specific RHOA G17V PCR for Diagnosis, Staging and Minimal Residual Disease Assessment in Angioimmunoblastic T-Cell Lymphoma

2018 ◽  
Vol 5 (1) ◽  
pp. 6-15
Author(s):  
Yu. Sidorova ◽  
Keyword(s):  
T Cell ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Cell Lymphoma ◽  
High Sensitivity ◽  
T Cell Lymphoma ◽  
Disease Assessment ◽  
Angioimmunoblastic T Cell Lymphoma ◽  
Allele Specific ◽  
Minimal Residual

scholarly journals Detection of the G17V RHOA Mutation in Angioimmunoblastic T-Cell Lymphoma and Related Lymphomas Using Quantitative Allele-Specific PCR

PLoS ONE ◽  
2014 ◽  
Vol 9 (10) ◽  
pp. e109714 ◽  
Author(s):  
Rie Nakamoto-Matsubara ◽  
Mamiko Sakata-Yanagimoto ◽  
Terukazu Enami ◽  
Kenichi Yoshida ◽  
Shintaro Yanagimoto ◽  
...  
Keyword(s):  
T Cell ◽  
Cell Lymphoma ◽  
T Cell Lymphoma ◽  
Angioimmunoblastic T Cell Lymphoma ◽  
Specific Pcr ◽  
Allele Specific ◽  
Allele Specific Pcr

scholarly journals Molecular diagnosis angioimmunoblastic T-cell lymphoma

2019 ◽  
Vol 91 (7) ◽  
pp. 63-69
Author(s):  
N G Chernova ◽  
Y V Sidorova ◽  
S Y Smirnova ◽  
N V Ryzhikova ◽  
E E Nikulina ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Lymph Node ◽  
T Cell ◽  
Peripheral Blood ◽  
Cell Lymphoma ◽  
Cell Receptor ◽  
T Cell Lymphoma ◽  
Tissue Samples ◽  
Angioimmunoblastic T Cell Lymphoma ◽  
Allele Specific

Aim: to determine molecular diagnostics routine for different tissue samples in angioimmunoblastic T-cell lymphoma. Materials and methods. Molecular studies were performed for 84 primary AITL patients. The median age was 61 year (29-81); the male to female ratio was 48/36. T-cell and B-cell clonality was assessed by GeneScan analysis of rearranged T-cell receptor (TCRG, TCRB) and immunoglobulin heavy chain genes. For the quantitative determination of cells with RHOA G17V mutation real - time polymerase chain reaction (PCR) with allele - specific LNA modified primers was used. Results. In lymph nodes rearrangements of T-cell receptor genes were determined in 76 (90.5%) of 84 patients and were absent in 8 (9.5%) cases. Identification of the same clonal products of the TCRG and TCRB genes in the lymph node and in peripheral blood and/or bone marrow indicated the prevalence of the tumor process and was observed in 64.7% of patients. Clonal products in peripheral blood and/or bone marrow different from those in the lymph node indicated reactive cytotoxic lymphocyte population and were noted in 58.8% of AITL cases. Simultaneous detection of T- and B-cell clonality in the lymph node was observed in 20 (24.7%) of 81 patients. Cells with RHOA G17V mutation were detected in lymph node in 45 (54.9%) of 82 patients. The use of allele - specific PCR with LNA modified primers revealed presence of the tumor cells in peripheral blood in 100% and in bone marrow in 93.9% of patients with G17V RHOA mutation in the lymph nodes. Conclusion. The validity of different molecular assays performed on certain tissue samples for the diagnosis of angioimmunoblastic T-cell lymphoma has been evaluated. Quantitative allele - specific PCR assay for RHOA G17V mutation based on LNA modified primers possesses sufficient sensitivity for tumor process prevalence evaluation and minimal residual disease monitoring.

scholarly journals Assessment of Clonotypic Rearrangements and Minimal Residual Disease in Lymphoid Malignancies: A Large Cancer Center Experience Using clonoSEQ

2021 ◽  
Author(s):  
Mohammad O. Hussaini ◽  
Jaya Srivastava ◽  
Lik Wee Lee ◽  
Taiga Nishihori ◽  
Bijal D. Shah ◽  
...  
Keyword(s):  
T Cell ◽  
Minimal Residual Disease ◽  
Plasma Cell ◽  
Residual Disease ◽  
Cell Lymphoma ◽  
B Cell Lymphoma ◽  
Response Strategy ◽  
Cancer Center ◽  
Lymphoid Malignancies ◽  
Minimal Residual

Context.— Measurable (minimal) residual disease (MRD) is an independent prognostic factor for survival outcomes in patients with lymphoid and plasma cell malignancies and has been incorporated into consensus criteria regarding treatment response, strategy, and clinical trial endpoints. clonoSEQ (a next-generation sequencing [NGS]-MRD assay) uses multiplex polymerase chain reaction and NGS to identify clonotypic rearrangements at the immunoglobulin (Ig) H, IgK, IgL, T-cell receptor (TCR)-β, and TCR-γ loci, and translocated B-cell lymphoma 1/IgH and 2/IgH sequences for MRD assessment. Additionally, it can be used to confirm diagnoses of cutaneous T-cell lymphoma (CTCL). Objective.— To review the technical aspects of our experience using the clonoSEQ Assay in routine clinical practice. Design.— In this single-center experience, 390 patients with lymphoid and plasma cell malignancies were assessed with the NGS-MRD Assay at a central laboratory. Results.— Median time from arrival of the shipment to initiation of the assay (defined as captured in Adaptive's secure tracking system) was 2.1 hours. Overall, 317 patients had 1 or more samples submitted for sequence identification. Of these, 290 (91.5%) had trackable sequences identified. The median calibration rate of samples by malignancy (where n ≥ 10 samples, excluding CTCL samples) was 88.1%, across a variety of fresh and archived sample sources (177 of 201 samples). TCR-β and/or TCR-γ clonotypes were identified in 40 of 95 samples (42.1%) from 66 patients with suspected CTCL. Conclusions.— This NGS-MRD Assay is a valuable and sensitive tool for monitoring MRD in patients with plasma cell and lymphoid malignancies and assisting in the diagnosis of CTCL.

scholarly journals Monitoring minimal residual disease in canine lymphomas treated with modified L-COP or L-CHOP protocols

2018 ◽  
Vol 66 (1) ◽  
pp. 66-84
Author(s):  
Sirintra Sirivisoot ◽  
Patharakrit Teewasutrakul ◽  
Somporn Techangamsuwan ◽  
Sirikachorn Tangkawattana ◽  
Anudep Rungsipipat
Keyword(s):  
T Cell ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
Cell Lymphoma ◽  
Clinical Stage ◽  
Progression Free Survival ◽  
B Cell Lymphoma ◽  
Blood Leukocytes ◽  
Free Survival ◽  
Minimal Residual

Heteroduplex polymerase chain reaction for antigen receptor rearrangements (hPARR) was developed to monitor minimal residual disease (MRD) in canine B- and T-cell lymphomas treated with the modified L-COP or L-CHOP protocol. Thirty-five dogs were recruited in this study and their neoplastic lineages were determined by immunophenotyping with Pax5 and CD3. Peripheral blood leukocytes were collected prior to and during chemotherapy in weeks 4, 9 and 13 to detect MRD by hPARR. Twenty-eight dogs (80%) had B-cell lymphoma while seven dogs (20%) had T-cell lymphoma. A monoclonal band was detected in 11 cases that showed complete or partial remission before tumour relapse and no response to the current treatment without statistical difference in clinical outcomes; however, the treatment response had an association with the MRD result (P < 0.05). Modified L-CHOP prolonged median progression-free survival as compared to modified L-COP (215 days vs. 93 days; P < 0.05). Substage b had shorter progression-free survival than substage a (90 days vs. 215 days; P < 0.05). Clinical stage III affected median overall survival time when compared to clinical stages IV and V (432, 173 and 118 days, respectively; P < 0.05). hPARR could be used for screening refractory lymphoma together with lymph node measurement in routine clinical cases.

scholarly journals Harmonization of Flow Cytometric Minimal Residual Disease Assessment in Multiple Myeloma in Centers of Polish Myeloma Consortium

Diagnostics ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1872
Author(s):  
Agnieszka Krzywdzińska ◽  
Bartosz Puła ◽  
Anna Czyż ◽  
Beata Krzymieniewska ◽  
Jolanta Kiernicka-Parulska ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Clinical Trials ◽  
Minimal Residual Disease ◽  
Residual Disease ◽  
High Sensitivity ◽  
Disease Assessment ◽  
Reproducibility Study ◽  
Highly Sensitive ◽  
Data Files ◽  
Minimal Residual

Minimal residual disease (MRD) status is now considered as one of the most relevant prognostic factors in multiple myeloma (MM) while MRD negativity became an important endpoint in clinical trials. Here, we report the results of the first study evaluating the reproducibility of high-sensitivity flow cytometry MM MRD assessment in four laboratories in Poland. EuroFlow protocols for instrument setting standardization and sample preparation in MM MRD assessment were implemented in each laboratory. In the inter-laboratory reproducibility study, 12 bone marrow samples from MM patients were distributed and processed in participant laboratories. In the inter-operator concordance study, 13 raw data files from MM MRD measurements were analyzed by five independent operators. The inter-laboratory study showed high 95% overall concordance of results among laboratories. In the inter-operator study, 89% of MRD results reported were concordant, and the highest immunophenotype interpretation differences with regard to expression of CD27, CD45, CD81 were noticed. We confirmed the applicability and feasibility of the EuroFlow protocol as a highly sensitive method of MRD evaluation in MM. Results of our inter-center comparison study demonstrate that the standardization of MM MRD assessment protocols is highly desirable to improve quality and comparability of results within and between different clinical trials.

scholarly journals Minimal Residual Disease Assessment in CLL/SLL: Is It Ready for Routine Clinical Practice?

2021 ◽  
Vol 19 (11.5) ◽  
pp. 1343-1345
Author(s):  
Jennifer A. Woyach
Keyword(s):  
Minimal Residual Disease ◽  
Residual Disease ◽  
High Sensitivity ◽  
Lymphocytic Leukemia ◽  
Disease Assessment ◽  
Fixed Duration ◽  
Btk Inhibitor ◽  
Term Data ◽  
Minimal Residual

Minimal residual disease (MRD) in chronic lymphocytic leukemia (CLL) is defined as <1 CLL cell per 10,000 leukocytes (0.01%; <10−4). Flow cytometry and next-generation sequencing have demonstrated high sensitivity in MRD detection. MRD assessment may help to determine prognosis after fixed-duration regimens; this has been established in the contexts of chemoimmunotherapy and venetoclax/antibody combinations. In the short term, MRD status does not seem to inform prognosis in patients treated with a BTK inhibitor plus venetoclax-based regimens; however, long-term data will be needed to determine whether it is beneficial in this population. Numerous trials have demonstrated that MRD may be used to guide therapy. It is unclear whether using an MRD-guided treatment strategy is better than using fixed-duration therapy; ongoing and future studies are warranted.

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