Analytical Detection of Immunoglobulin Heavy Chain Gene Rearrangements in Gastric Lymphoid Infiltrates by Peak Area Analysis of the Melting Curve in the LightCycler System

Abstract

Because it is difficult to differentiate gastric mucosaassociated lymphoid tissue (MALT) lymphoma from chronic gastritis in gastric lymphoid infiltrates, molecular detection of monoclonality through immunoglobulin heavy chain (IgH) gene rearrangements is commonly performed. However, heterogeneity in the performance and results obtained from IgH gene rearrangements has been reported. To improve the accuracy in the diagnosis of gastric lymphoid infiltrates, we developed an analytical approach based on one-peak area analysis of the melting curve in the LightCycler System. Using a training-testing approach, the likelihood ratio method was selected to find a discriminative function of 4.64 in the training set (10 gastric MALT lymphomas and 10 chronic gastritis cases). This discriminative function was validated in the testing set (five gastric MALT lymphomas, six abnormal lymphocytic infiltrates with subsequently demonstrated gastric MALT lymphomas, and six cases of chronic gastritis). All but one case of gastric MALT lymphoma, as well as abnormal lymphocytic infiltrates, clustered under 4.64, and all chronic gastritis cases clustered above 4.64. These results were validated by conventional electrophoreses confirming one or two sharp bands in cases of gastric MALT lymphomas and a smear of multiple bands in cases of chronic gastritis. Analytical detection of IgH gene rearrangement in gastric lymphoid infiltrates by one-peak area analysis correctly distinguishes gastric MALT lymphomas from chronic gastritis, even in cases with diagnosis of abnormal lym-

Rapid detection of clonal expansion of T‐cell receptor‐beta gene in patients with HBV using the real‐time PCR with DNA melting curve analysis

“…Compared to other methods for detecting TCRBV gene families, RQ‐PCR has high sensitivity and repeatability. Moreover, RQ‐PCR data can be analyzed directly, without agarose gel electrophoresis to detect the PCR product, which saves experimental time, increases experimental efficiency and reduces the chance of cross contamination during operation 25 …”

“…Moreover, RQ-PCR data can be analyzed directly, without agarose gel electrophoresis to detect the PCR product, which saves experimental time, increases experimental efficiency and reduces the chance of cross contamination during operation. 25 In our study, the experimental data were analyzed post-PCR using the Option Monitor 3.0 software (Bio-rad, Hercules, CA) to obtain the -dF/dT, and the latter was plotted against the Tm to obtain the peak pattern. There are 24 TCRBV gene families composing the GMSP of a sample, and every GMSP consists of a series of melting curve peaks.…”

Melting curve analysis in canine lymphoma by calculating maximum fluorescence decrease

“…Efforts to standardize the interpretation of melting curves have already been made in human medicine. Retamales et al used the LightCycler data analysis software to calculate one peak area under the curve and then used the likelihood ratio to distinguish between polyclonal and monoclonal samples …”

Reprimo as a Potential Biomarker for Early Detection in Gastric Cancer

“…To identified the DNA methylation patterns of 24 genes, we used a testing-validation approach (12). As a testing set, we selected 32 surgically resected gastrectomy specimens with diagnoses of gastric cancer according to the WHO (13).…”