TY - JOUR
T1 - Clonality analysis of B-cell lymphoma in fresh-frozen and paraffin-embedded tissues
T2 - the effects of variable polymerase chain reaction parameters.
AU - Chen, Y. T.
AU - Whitney, K. D.
AU - Chen, Y.
PY - 1994/5
Y1 - 1994/5
N2 - To investigate the sensitivity of polymerase chain reaction (PCR)-based detection of B-cell monoclonality and the effects of several variables, we analyzed 119 cases of B-cell lymphomas with proven IgH gene rearrangements, testing fresh-frozen and formalin-fixed materials in parallel. Using fresh-frozen tissue, 83 cases (70%) were positive with one-step PCR and high-stringency annealing. Two groups of false-negative cases were identified, Group I (16 cases) showing no PCR products, and Group II (20 cases) showing polyclonal smear patterns. Seminested PCR and/or lowered annealing stringency revealed nine additional positive cases in Group I but none in Group II. Amplification with bcl-2/JH primers resulted in five more positive cases. The overall positive rate in the fresh-frozen category was 81% (97 of 119). Parallel analysis was performed on formalin-fixed, paraffin-embedded material from 61 cases, and a high concordance rate (89%) was observed. The results indicated that fresh and formalin-fixed specimens are comparable in this PCR-based assay, and that the two groups of false-negative results can be accounted for by different reasons. In addition, we reviewed the current literature, discussed the diagnostic applications of this technique, and listed the core elements of a proposed PCR protocol that should be suitable for most laboratories.
AB - To investigate the sensitivity of polymerase chain reaction (PCR)-based detection of B-cell monoclonality and the effects of several variables, we analyzed 119 cases of B-cell lymphomas with proven IgH gene rearrangements, testing fresh-frozen and formalin-fixed materials in parallel. Using fresh-frozen tissue, 83 cases (70%) were positive with one-step PCR and high-stringency annealing. Two groups of false-negative cases were identified, Group I (16 cases) showing no PCR products, and Group II (20 cases) showing polyclonal smear patterns. Seminested PCR and/or lowered annealing stringency revealed nine additional positive cases in Group I but none in Group II. Amplification with bcl-2/JH primers resulted in five more positive cases. The overall positive rate in the fresh-frozen category was 81% (97 of 119). Parallel analysis was performed on formalin-fixed, paraffin-embedded material from 61 cases, and a high concordance rate (89%) was observed. The results indicated that fresh and formalin-fixed specimens are comparable in this PCR-based assay, and that the two groups of false-negative results can be accounted for by different reasons. In addition, we reviewed the current literature, discussed the diagnostic applications of this technique, and listed the core elements of a proposed PCR protocol that should be suitable for most laboratories.
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M3 - Article
C2 - 8066071
AN - SCOPUS:0028433238
SN - 0893-3952
VL - 7
SP - 429
EP - 434
JO - Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc
JF - Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc
IS - 4
ER -