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<p><b>OBJECTIVE</b>To explore the feasibility of detecting lymphoma with the application of BIOMED-2 standardized immunoglobulin/T cell receptor (IG/TCR) gene rearrangement system in formalin fixed paraffin-embedded (FFPE) tissue samples, and to discuss the relationship between the longest amplification fragment of extracted DNA and positive detection rate of different IGH V-J primers.</p><p><b>METHODS</b>DNA was extracted from 50 cases of FFPE tissue samples. Multiplex-PCR amplifications were performed and then the IG/TCR gene rearrangements were analyzed using BIOMED-2 standardized clonality analysis system.</p><p><b>RESULTS</b>(1)When the DNA concentration was diluted to 50-100 ng/μl from 100-500 ng/μl, the proportion of the longest amplification fragment (300-400 bp) of DNA was improved from 10.0% to 90.0% in 30 cases of diffuse large B cell lymphoma (DLBCL) wax roll samples (P<0.01). The positive rate of IGH+IGK was increased from 46.7% to 83.3%, the difference was statistically significant (P=0.006). The lengths of the longest amplification fragments of DNA were all longer than 300 bp in the paraffin section samples of DLBCL. The positive rate of IGH+IGK of these samples was 96.7%. The difference of the positive rate of IGH+IGK between the wax roll samples and the paraffin section samples had no statistical significance (P=0.195). (2)When the concentration of DNA was high, most of the longest amplification fragments of extracted DNA were 100 bp or 200 bp, and the detection rate of short fragment IGH FR3 was more stable than that of long fragment IGH FR1. (3)The clonality analysis of TCRG+TCRB in all 13 cases of peripheral T cell lymphoma samples showed positive results, while no positive IG/TCR clones were found in 7 cases of reactive lymphoid tissue hyperplasia in control group.</p><p><b>CONCLUSION</b>Dilution of DNA is the only method to improve not only the proportion of longest fragment amplification but also the detection rate of clonality. The detection rate of IGH FR3 would not be affected by the concentration of DNA. The application of BIOMED-2 standardized IG/TCR gene rearrangement system in FFPE tissue samples plays an important role in the lymphoma diagnosis.</p>
Assuntos
Humanos , Rearranjo Gênico do Linfócito T , Linfoma , Diagnóstico , Genética , Inclusão em Parafina , Recombinação V(D)JRESUMO
Objective:The occurrence of numerous tumors, particularly classical Hodgkin's lymphoma (CHL), is related with Ep-stein-Barr virus (EBV) infection. However, the incidence of CHL and its association with EBV varies significantly with ethnicity, geo-graphic location, sex, and age. This study investigated the association of EBV infection with CHL in Northern Chinese Han population. Methods:EBV-encoded small RNA (EBER) was detected in 136 cases of CHL through in situ hybridization. Results:A total of 37 cas-es were EBER positive (28%). The mixed cellularity (MC) subtype had the highest positive EBER rate of 49%(23/47;P<0.001), fol-lowed by lymphocyte-rich subtype with 30%(3/10), nodular sclerosis (NS) subtype with 14%(10/73), and 1ymphocyte depletion with 0%(0/2). Our study identified a single age distribution in the third decade. Moreover, NS subtype showed an evident single peak in the third decade. However, MC subtype had a lower peak in the fifth decade. The incidence of EBER showed a bimodal age distribution with two peaks in the first and fifth decades (21.6%and 24.3%, respectively). Conclusion:CHL in Northern Chinese Han population was associated with EBV infection, particularly the MC subtype.
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EB virus (EBV) is a ubiquitous y-herpes virus,which asymptomatically infects greater than 90 ％ of the world's population.EBV is closely related to development of several kinds of lymphomas that include B-,T-and NK-cell derived.Subsequent studies have proven that EBV gene expression product plays an activating and/or promoting role on lymphomagenesis,and paves the way for EBV associated lymphomas immunotherapy.This review focuses on the EBV associated lymphomas in 2008 WHO classification of tumours of haematopoietic and lymphoma tissues.
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Anaplastic lymphoma kinase-positive (ALK+) diffuse large B-cell lymphoma (DLBCL) is a rare subtype of DLBCL. ALK+DLBCL has a characteristic immunoblastic/plasmablastic morphology, a distinct immunophenotypic profile, and recurrent cytoge-netic/molecular genetic abnormalities. The occurrence of this type of lymphoma has been reported in both adult and pediatric popula-tions. Although rare, this new entity should be recognized because most cases follow an aggressive clinical course with a poor progno-sis. The response of ALK+DLBCL to conventional chemotherapy is poor. The recently discovered small molecule ALK inhibitor may provide a potential therapeutic option for patients with this disease.