Failure of PCR Tests

In contrast to general clinical molecular PCR tests, apparent failure of the reaction in T-cell clonality assays can be due to a paucity of T cells rather than poor-quality DNA or the presence of PCR inhibitors such as heparin. To rule out an intrinsic problem with the amplification itself, the quality of the DNA is assessed by amplification of an unrelated control gene. False-negative results can largely be avoided by optimal selection of primers, as well as amplification of a control sequence for each sample, in addition to clonality testing. The detection rate for TCRG PCR testing also depends on the specimen type and quality used for DNA extraction. A significantly lower number of true positives may be achieved using paraffin-embedded tissue, and laboratories often validate the sensitivity of the assay specifically for paraffin-embedded tissue specimens.

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