ADDITIONAL TCRV-BETA PRIMERS AND MINOR METHOD MODIFICATIONS IMPROVE DETECTION OF CLONAL T-CELL POPULATIONS BY RT-PCR

The TCRV beta RT-PCR method for detection of clonal populations of T c ells which we described previously could not detect clones that used c ertain variable (V) beta region families. V beta 2, 4, 8.3, and 18 had insufficient homology with the original consensus V region primer. Tw o new primers have been designed which work well and are able to ampli fy from V beta families previously undetectable by this RT-PCR. In add ition, minor alterations to the cDNA synthesis and gel analysis of the make the results even interpret. All the Diversity/Joining (D/J) regi on primer combinations except for D2/J2 have been omitted, and termina ting the reverse transcription by heating prior to PCR greatly improve s amplification with these primers. Use of 8% and/or 10% polyacrylamid e gels increases clarity. Inclusion of the modifications described wil l reduce false reporting of patients as having a polyclonal T-cell pop ulation.