Fj. Child et al., Molecular analysis of the immunoglobulin heavy chain gene in the diagnosisof primary cutaneous B cell lymphoma, J INVES DER, 117(4), 2001, pp. 984-989
The diagnosis of primary cutaneous B cell lymphoma can be difficult on the
basis of histologic and immunophenotypic features alone. Previous polymeras
e chain reaction studies for detection of a clonal population in nodal B ce
ll lymphomas have employed different primer pairs with detection sensitivit
ies varying between 34% and 94% but there have been no comprehensive studie
s of primary cutaneous B cell lymphoma. We compared the sensitivity of diff
erent sets of consensus primers to amplify the CDR3 VDJ region of the immun
oglobulin heavy chain gene in combination with an immunoglobulin heavy chai
n joining region consensus primer to detect a monoclonal population in 39 c
ases of primary cutaneous B cell lymphoma. Radiolabeled products were analy
zed with denaturing 6% polyacrylamide gel electrophoresis. Sequence analysi
s was used to confirm amplification of clonal immunoglobulin heavy chain ge
ne rearrangements and to establish whether somatic hypermutation can interf
ere with primer binding. Clonal immuno globulin heavy chain gene rearrangem
ents were demonstrated in 79% of cases (74% with leader sequences, 64% with
FR1, and 45% with FR3 primers). Somatic hypermutation at primer binding si
tes was confirmed in cases where a false negative result was obtained with
the FR3 primer. Although monoplex polymerase chain reaction amplification u
sing the leader sequence primers is the most sensitive method for detecting
a clonal population, six primers are required in six different reactions.
Our findings suggest initial analysis with the FR3 primer and subsequent an
alysis using leader sequences in negative cases. Our data indicate that the
FR3 consensus primer alone is not sufficient for a comprehensive analysis
of primary cutaneous B cell lymphoma.