K. Olsson et al., Real-time t(11;14) and t(14;18) PCR assays provide sensitive and quantitative assessments of minimal residual disease (MRD), LEUKEMIA, 13(11), 1999, pp. 1833-1842
Non-Hodgkin's lymphoma (NHL) arises as a clonal transformation of normal B
and T cell differentiation and is often characterized by a higher incidence
of specific chromosomal translocations, We have developed real-time TaqMan
PCR assays directed toward two of these tumor-associated DNA markers, the
t(14;18)(q32;q21.3) at the major breakpoint region of the bcl-2 gene and th
e t(11;14)(q13;q32) at the bcl-1 major translocation cluster. During analys
is of serial dilutions of t(14;18)positive DNA, the t(14;18) real-time PCR
was at least as sensitive as nested PCR and demonstrated enhanced quantitat
ive potential, Moreover, in a blinded comparison of the t(14;18) real-time
PCR and a clinically validated nested PCR protocol using 134 cell line and
patient DNA samples, the real-time PCR detected the translocation in 30.0%
more cases than nested PCR. Both the t(14;18) and t(11;14) real-time PCR as
says were used to quantitate minimal residual disease (MRD) in an NHL clini
cal trial assessing the safety and efficacy of a tumor-purging protocol in
autologous stem cell transplantation. The assays were also used to evaluate
disease depletion in an ex vivo tumor spiking model in which normal periph
eral blood was spiked with tumor cell lines and processed according to the
clinical purging method. PCR data from both the clinical trial and the ex v
ivo model demonstrated a 4 to 6 log reduction in tumor cells during CD34(+)
and CD34(+) Thy-1(+) enrichment. Because the t(14;18) and t(11;14) real-ti
me PCR assays are very sensitive, quantitative, rapid, and require no post-
PCR manipulation, they may serve as practical alternatives to nested PCR.