Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements

Abstract

Detecting clonal T-cell receptor (TCR)-␥ gene rearrangements (GRs) is an important adjunct test fordiagnosing T-cell lymphoma. We compared a recently described assay (BIOMED-2 protocol) , which targets multiple variable (V) gene segments in two polymerase chain reaction (PCR) reactions (multi-V) , with a frequently referenced assay that targets a single V gene segment in four separate PCR reactions (mono-V). A total of 144 consecutive clinical DNA samples were prospectively tested for T-cell clonality by PCR using laboratory-developed mono-V and commercial multi-V primer sets for TCR-␥ GR. The combination of TCR-␤ , mono-V TCR-␥ and multi-V TCR-␥ detected more clonal cases (68/144 , 47%) than any individual PCR assay. We detected clonal TCR-␤ GR in 47/68 (69%) cases. Using either mono-V or multi-V TCR-␥ primers , the sensitivities for detecting clonality were 52/68 (76%) or 51/68 (75%). Using both mono-V and multi-V TCR-␥ primers improved the sensitivity for detecting clonality , 60/68 (88%). Combining either mono-V or multi-V TCR-␥ primers with TCR-␤ primers also improved the sensitivity , 64/68 (94%). Significantly , TCR-␥ V11 GRs could only be detected using the mono-V-PCR primers. We conclude that using more than one T-cell PCR assay can enhance the overall sensitivity for detecting T-cell clonality.

A parallel comparison of T-cell clonality assessment between an in-house PCR assay and the BIOMED-2 assay leading to an efficient and cost-effective strategy

“…Performing other BIOMED-2 reactions is needed only for the remaining 9% of cases, of which 3% are likely to show a clonal TCRB and/or TCRD gene rearrangement. Such an overall level of T-cell clonality detection rate compares well with the 94% detection rate reported in the follow-up study of the BIOMED-2 group13 and the 93–96% detection rate reported in the independent studies using BIOMED-2 assays combined with laboratory developed assays on fresh-frozen materials 30 31. Thus, approximately 5% of false negative results are expected in T-cell malignancies with adequate DNA quality.…”

Lymph Node Fine-Needle Cytology: Beyond Flow Cytometry

“…Clonality analysis has been defined in the EuroClonality/BIOMED-2 consortium by the standardization of multiplex PCR assays for nearly all Ig/TCR targets (Igκ and TCRβ, incomplete IGH D-J and TCRB D-J rearrangements). These tests collectively show a high rate of detection in the most common B- and T-cell lymphomas [41,42,43,48,49]. EuroClonality/BIOMED-2 have also produced guidelines for the interpretation and for reporting results in suspected lymphoma [37].…”

“…Ig and TCR antigen receptor gene rearrangements are the mostly applied targets for clonality testing in NHL [42,43,48,49,50]. Ig and TCR rearrangements occur in the earliest stages of B and T cell development by the random joining of many V, (D) and J genes in a unique V(D)J exon that encodes the final antigen-binding moiety of the Ig or TCR chain [51].…”

Specific TCR gene rearrangements in mycosis fungoides: does advanced clinical stage show a preference?

“…TCR studies were performed at the time of diagnosis on formalin-fixed paraffin embedded (FFPE) tissue as previously described 14. Briefly, 20–30 sections of the FFPE tissue were cut at a thickness of 5 μm.…”