Dataset Information

Identification of the tumor cells in peripheral T-cell lymphomas by combined polymerase chain reaction-based T-cell receptor beta spectrotyping and immunohistological detection with T-cell receptor beta chain variable region segment-specific antibodies.

ABSTRACT: Most nodal peripheral T-cell lymphomas (PTCL) originate from alphabeta-T cells, and they often contain reactive T cells that may hamper immunophenotyping. To specifically identify the neoplastic population in immunohistochemically stained slides, we assessed the heterogeneity of the T-cell receptor beta chain variable region (TCRVbeta). This region contains 65 gene segments, of which only one is expressed after rearrangement. To investigate PTCL, we developed a polymerase chain reaction assay to define the clonally rearranged TCRVbeta segment. Detecting the corresponding epitope with segment-specific antibodies enabled identification of tumor cells among the T cells. The TCRVbeta segment of the tumor cells was defined in 13 of 13 PTCL not otherwise specified and 11 of 13 angioimmunoblastic T-cell lymphomas. Antibodies corresponding to the respective TCRVbeta segment of the tumor were available for seven cases from each group. After applying these antibodies in combination with antibodies against CD3, CD5, CD4, CD8, and cytotoxic molecules, double stains were evaluated by confocal laser scanning microscopy. In 9 of 14 cases, less than 50% of T cells expressed the clonally rearranged TCRVbeta segment. Phenotypes defined in double stains differed from those obtained by conventional immunohistochemistry in 11 of 14 cases. The combination of TCRVbeta polymerase chain reaction and immunohistochemistry may facilitate more reliable detection and characterization of tumor cells in PTCL.

SUBMITTER: Geissinger E

PROVIDER: S-EPMC1885558 | biostudies-literature | 2005 Oct

REPOSITORIES: biostudies-literature

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Identification of the tumor cells in peripheral T-cell lymphomas by combined polymerase chain reaction-based T-cell receptor beta spectrotyping and immunohistological detection with T-cell receptor beta chain variable region segment-specific antibodies.

Geissinger Eva E Bonzheim Irina I Krenács László L Roth Sabine S Ströbel Philipp P Ott German G Reimer Peter P Wilhelm Martin M Müller-Hermelink Hans Konrad HK Rüdiger Thomas T

The Journal of molecular diagnostics : JMD 20051001 4

Most nodal peripheral T-cell lymphomas (PTCL) originate from alphabeta-T cells, and they often contain reactive T cells that may hamper immunophenotyping. To specifically identify the neoplastic population in immunohistochemically stained slides, we assessed the heterogeneity of the T-cell receptor beta chain variable region (TCRVbeta). This region contains 65 gene segments, of which only one is expressed after rearrangement. To investigate PTCL, we developed a polymerase chain reaction assay to ...[more]

PMID: 16237215

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Project description:Background and aimDifferent polymerase chain reaction (PCR) techniques have and are still being used for the direct detection of Brucella DNA in serum samples of different animal species and humans without being validated or properly validated, resulting in discrepancies. Thus, this study aimed to evaluate the diagnostic performance of the TaqMan Real-Time-PCR (RT-PCR) targeting the bcsp31 gene versus conventional PCR for the accurate diagnosis of brucellosis at the genus level in cattle sera.Materials and methodsOne hundred and eighty-four serum samples were collected from bacteriologically positive and negative cows with ages ranging from 1 to 5 years old at some infected private farms in the Nile Delta under quarantine measures as well as brucellosis free farms. These samples were classified into four groups after serological diagnosis and investigated by TaqMan RT-PCR and conventional PCR targeting the IS711 gene for Brucella DNA detection. The diagnostic performance characteristics of both PCR techniques were estimated considering the bacteriological results as a gold standard.ResultsTaqMan RT-PCR revealed superiority over conventional PCR; it was able to detect Brucella DNA in 95% (67/70) and 89% (25/28) of the cattle sera samples belonging to Group 1 (serologically and bacteriologically positive) and Group 2 (serologically negative but bacteriologically positive), respectively. On evaluating the diagnostic performance, TaqMan RT-PCR showed superior diagnostic sensitivity (93.9%), diagnostic specificity (88.4%), performance index (182.3), almost perfect kappa agreement (0.825±0.042), strong positive correlation (r=0.826), high accuracy based on the receiver operating characteristic (ROC) curve, and area under the ROC curve (0.911) at p<0.05 and CI of 95%.ConclusionA cattle serum sample is not the metric of choice for targeting Brucella genomic DNA by conventional PCR. The time-saving and rapid TaqMan RT-PCR method revealed a better diagnostic performance in the detection of Brucella DNA in cattle sera. Such performance offered by TaqMan RT-PCR may be considered a step toward the possibility of using such technology in the direct differentiation between Brucella-infected and -vaccinated cattle immunized by smooth vaccines from cattle sera using primers specific for such vaccines.

Dataset Information

Identification of the tumor cells in peripheral T-cell lymphomas by combined polymerase chain reaction-based T-cell receptor beta spectrotyping and immunohistological detection with T-cell receptor beta chain variable region segment-specific antibodies.

Publications

Identification of the tumor cells in peripheral T-cell lymphomas by combined polymerase chain reaction-based T-cell receptor beta spectrotyping and immunohistological detection with T-cell receptor beta chain variable region segment-specific antibodies.

Similar Datasets

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