Comparison of different techniques to detect MRD in MCL
| Method . | MFC . | Consensus PCR . | Nested PCR . | RQ-PCR . | HTS . |
|---|---|---|---|---|---|
| Target | Immunophenotype | IGH rearrangement or t(11;14) | IGH rearrangement or t(11;14) | IGH rearrangement or t(11;14) | IGH rearrangement |
| Reproducible detection limit | 10−3 to 10−4 (4-color MFC) | IGH: 10−2 to 10−3 | 10−5 | 10−5 | Up to 10−6 (dependent on DNA amount) |
| 10−4 (8-color MFC) | t(11;14): 10−4 | ||||
| Level of information | Quantitative (above detection limit) | Qualitative | Qualitative | Quantitative (above limit of quantification) | Quantitative (above detection limit) |
| Patient-specific PCR primer needed | Not applicable | No | Depending on approach | Yes | No |
| Patient applicability in advanced-stage MCL | >85% | >95% | >85% | >85% | No data yet |
| Expertise | High for 6- to 8-color MFC | Lower | High | High | High |
| Standardization | No | No | No | Yes | No |
| Turnaround time | 3-4 h | 3-4 h | Dependent on method; mostly 3-4 h | 2 wk | 1 wk |
| Advantages | Rapid quantification | Rapid and robust | No establishment of serial dilution for quantification needed | High sensitivity; greatest interlaboratory reproducibility; regular quality control rounds | High sensitivity; independent of patient-specific primers; additional information on background B-cell repertoire |
| Disadvantages | Low sensitivity; expertise needed for evaluation; not standardized | Low sensitivity; unspecific amplification possible; no quantification | Not quantitative; not standardized | ASO primer design necessary; time-consuming workflow | Super multiplex PCR (disproportional target amplification possible); discrimination from normal cell background (requires about 5% tumor cell infiltration); complex bioinformatic evaluation |
| Method . | MFC . | Consensus PCR . | Nested PCR . | RQ-PCR . | HTS . |
|---|---|---|---|---|---|
| Target | Immunophenotype | IGH rearrangement or t(11;14) | IGH rearrangement or t(11;14) | IGH rearrangement or t(11;14) | IGH rearrangement |
| Reproducible detection limit | 10−3 to 10−4 (4-color MFC) | IGH: 10−2 to 10−3 | 10−5 | 10−5 | Up to 10−6 (dependent on DNA amount) |
| 10−4 (8-color MFC) | t(11;14): 10−4 | ||||
| Level of information | Quantitative (above detection limit) | Qualitative | Qualitative | Quantitative (above limit of quantification) | Quantitative (above detection limit) |
| Patient-specific PCR primer needed | Not applicable | No | Depending on approach | Yes | No |
| Patient applicability in advanced-stage MCL | >85% | >95% | >85% | >85% | No data yet |
| Expertise | High for 6- to 8-color MFC | Lower | High | High | High |
| Standardization | No | No | No | Yes | No |
| Turnaround time | 3-4 h | 3-4 h | Dependent on method; mostly 3-4 h | 2 wk | 1 wk |
| Advantages | Rapid quantification | Rapid and robust | No establishment of serial dilution for quantification needed | High sensitivity; greatest interlaboratory reproducibility; regular quality control rounds | High sensitivity; independent of patient-specific primers; additional information on background B-cell repertoire |
| Disadvantages | Low sensitivity; expertise needed for evaluation; not standardized | Low sensitivity; unspecific amplification possible; no quantification | Not quantitative; not standardized | ASO primer design necessary; time-consuming workflow | Super multiplex PCR (disproportional target amplification possible); discrimination from normal cell background (requires about 5% tumor cell infiltration); complex bioinformatic evaluation |
IGH, immunoglobulin heavy chain gene.