Detection of concurrent/recurrent non-hodgkin's lymphoma in effusions by PCR

Murphy, Michael J.; Signoretti, Sabina; Nasser, Imad; Sherburne, Bradford; Loda, Massimo

doi:10.1016/s0046-8177(99)90069-2

Cited by 11 publications

(11 citation statements)

References 26 publications

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“…A monoclonal pattern was demonstrated in all lymphomatous effusions by IgH gene configuration analysis. In case 2, the monoclonal IgH pattern was also confirmed after using the semi‐nested amplification protocol of the CDR3 region31 and PEL‐derived clonotypic primers. Heterogeneously sized IgH multiple rearrangements without reinforced single bands, corresponding to polyclonal B‐cells, were amplified in nodal and spleen MCD lesions removed before the diagnosis of PEL (cases 1, 2, and 4) or obtained at autopsy (case 3), and in several organs containing atypical lymphoid infiltrates (cases 1, 2, and 3).…”

Section: Resultsmentioning

confidence: 85%

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Primary effusion lymphoma in HIV-infected patients with multicentric Castleman's disease

et al. 2001

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Multicentric Castleman's disease (MCD) and primary effusion lymphoma (PEL) are two B-cell lymphoproliferative diseases associated with Kaposi's sarcoma-associated herpes virus/human herpesvirus-8 (KSHV/HHV-8). Although MCD is considered a prelymphoma state, it is not known whether a pathogenetic link exists between MCD and PEL. This paper reports the clinico-pathological features of four cases of PEL (two pericardial, one pleural, and one peritoneal) developing in the context of HIV-associated MCD. Effusions, lymph nodes, spleen, and additional tissues from three autopsies were examined for morphology/immunophenotype, search for HHV-8 DNA, and assessment of immunoglobulin heavy chain gene (IgH) configuration using polymerase chain reaction (PCR)-based techniques. MCD and PEL samples contained HHV-8 DNA. Clonal IgH rearrangements were detected only in PEL, whereas MCD tissues were polyclonal. Light-chain immunostaining confirmed B-cell clonality in PEL (two lambda, one kappa, one not tested) and polyclonality in MCD. The autopsies revealed different morphological variants of visceral KS and multi-organ atypical infiltrates exhibiting immunoblastic/plasmablastic features reminiscent of PEL morphology, with a restriction of lambda-positive cells. In two cases, using microdissection and IgH PCR analysis, multiple/discrete bands were found in the infiltrates, compatible with polyclonality/oligoclonality. The case showing an oligoclonal IgH ladder contained a rearrangement of identical junctional size to the PEL clone; however, further analysis with PEL-derived clonotypic primers and sequencing of PCR products showed no amplification and nucleotide diversity, respectively, indicating that the two B-cell populations examined were clonally unrelated. These data show that MCD and PEL may co-exist in HIV-infected patients, suggesting a relevant association between these two HHV-8-related disorders. Although a definite clonal relationship between MCD and PEL was not demonstrated, it is hypothesized that in some MCD cases, within expanded polyclonal B-cell populations secondary to HHV-8 infection, clonal expansions may occur that localize into a body cavity, i.e. PEL.

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Section: Resultsmentioning

confidence: 85%

“…Each sample contained approximately 100–500 cells. The DNA extraction, semi‐nested protocol, and primers for PCR amplification of the IgH CDR3 region have been reported elsewhere31. DNA from each sample was amplified at least twice.…”

Section: Methodsmentioning

confidence: 99%

Primary effusion lymphoma in HIV-infected patients with multicentric Castleman's disease

et al. 2001

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“…30 -32 However, if diagnostic doubt remains over lymphoma versus inflammation, we can confirm that DNA extracted from formalin-fixed, paraffin-embedded tissues can be amplified successfully, permitting the analysis of clonality in archival material. 22,[33][34][35] Standard DNA extraction methods are time consuming and involve multiple tube transfers in which cells and DNA may be lost. For the purpose of application in ocular samples, in which the sample volume is small, Qiagen DNA Qiamp mini kit was used to reduce the potential loss of cells and DNA in transfer between tubes.…”

Section: Discussionmentioning

confidence: 99%

“…2,24,32,35 Diagnostic accuracy can be improved by using ancillary immunohistochemistry if sufficient material is available. For example, B-cell monoclonality may be inferred by demonstrating monotypic immunoglobulin (Ig) light chain expression using immunohistochemistry.…”

Section: Discussionmentioning

confidence: 99%

Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples

Lobo

Okhravi

Adamson

et al. 2007

The Journal of Molecular Diagnostics

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To determine the usefulness of polymerase chain reaction (PCR) analyses in the diagnosis of lymphoid infiltrate cells in ocular samples, PCR was performed using oligonucleotide primers specific for immunoglobulin heavy chain rearrangement at framework 2, framework 3, and t(14;18) translocation of the bcl-2 gene. These were used to successfully generate amplicons of 220 to 230 bp, 110 to 120 bp, and 175 to 200 bp, respectively. After PCR amplification, primers directed against the t(14;18) detected 10 pg of B-cell lymphoma DNA. PCR against Fr2 and Fr3 IgH rearrangement detected 10 fg and 10 pg in the seminested PCR, respectively. Conventional pathological methods were highly accurate at establishing the correct final diagnosis in formalin-fixed, paraffin-embedded samples but were much less sensitive and predictive in cytological specimens of intraocular fluid. A combination of the three PCR reactions was an equally successful diagnostic approach on paraffin-embedded samples, whereas single PCR reactions did not significantly improve diagnosis over histopathological diagnostic techniques. Thus, a combination of PCR reactions is useful in the detection of B-cell monoclonality, aids the differentiation between lymphomatous and inflammatory infiltrates, and is more powerful as a diagnostic method than single PCR or conventional cytopathology for lymphoid infiltrates in ocular fluid aspirates.

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“…During the last 10 yr, clonality analysis using molecular genetics has been increasingly used in surgical pathology and hematopathology 10 but less in cytology. [11][12][13][14][15][16][17] Molecular genetic analysis seems to be an attractive method for determining clonality, since only a small quantity of material is required.…”

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confidence: 99%

Application of molecular genetics to the diagnosis of lymphoid‐rich effusions: Study of 95 cases with concomitant immunophenotyping

Mihaescu

Gebhard

Chaubert

et al. 2002

Diagnostic Cytopathology

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The cytological differentiation between reactive lymphocytosis and malignant lymphoma in serous effusions is often difficult. The present study was designed to evaluate the potential contribution of molecular genetic clonality analysis to a solution to this problem. We examined the cytological specimens of 95 consecutive patients collected during a 4-yr period, including 74 pleural, 20 peritoneal, and one pericardial fluids. Cytological diagnosis in the 95 lymphocyte-rich effusions was positive for lymphoma in 20 cases, suspicious for lymphoma in 26 cases, and negative in 49 cases. The analysis by ICC was not carried out, inconclusive, or noninterpretable in 25 cases. In five cases molecular genetic analysis was hampered by technical problems. By immunocytochemistry, eight additional cases of lymphoma were detected and lineage classification was achieved in 15 of the 20 cytologically positive effusions. PCR and Southern blot analysis were used to assess B- and T-cell clonality. Monoclonality was found in 40 (42%) of the 95 effusions analyzed. One-third of the effusions with a monoclonal B-cell gene rearrangement were detected by Southern blot analysis but not by the PCR performed in parallel. The results of molecular genetic analysis were corroborated by histological findings and/or clinical evolution in 15 cases. Our results indicate that molecular genetic analysis is a useful tool in the analysis of lymphocyte-rich serous effusions.

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Detection of concurrent/recurrent non-hodgkin's lymphoma in effusions by PCR

Cited by 11 publications

References 26 publications

Primary effusion lymphoma in HIV-infected patients with multicentric Castleman's disease

Primary effusion lymphoma in HIV-infected patients with multicentric Castleman's disease

Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples

Application of molecular genetics to the diagnosis of lymphoid‐rich effusions: Study of 95 cases with concomitant immunophenotyping

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