Sa. Weston et al., Assessment of male CVID patients for mutations in the Btk gene: how many have been misdiagnosed?, CLIN EXP IM, 124(3), 2001, pp. 465-469
The presentation of hypogammaglobulinaemia in young males without a family
history of immunodeficiency can pose a diagnostic problem. In the past, the
presence of B-cells has suggested a diagnosis of common variable immunodef
iciency (CVID), although genotypic analysis has now clarified that individu
als with B cells may have mutations in their Btk gene. In order to address
the issue of how many male individuals with a clinical diagnosis of CVID do
in fact have mutations in the Btk gene, we analysed a group of 24 male pat
ients. Single-strand conformation polymorphism (SSCP) analysis was used to
screen the patient cohort for mutations in the Btk gene. Given the size of
the Btk gene, the number of patients in the cohort and the amount of availa
ble DNA, multiplex PCR reactions were utilized to span the 19 exons and pro
moter region of the gene. Where abnormal migration patterns were observed w
ith multiplex PCR reactions, in nine of the 24 patients, the individual Btk
gene fragments were re-amplified and analysed again by SSCP. Following thi
s analysis, four patients continued to demonstrate abnormal SSCP migration
patterns. However, direct sequencing of the relevant Btk gene fragments for
these four CVID patients revealed a mutation in only one patient. The muta
tion was the previously described polymorphism at position 2031 of Btk gene
within exon 18. These results indicate that caution should be taken with t
he application of SSCP analysis to mutation detection. While it has a role
to play in screening large patient cohorts, direct sequencing is a necessar
y adjunct to such analysis. Finally, the clinical diagnosis of CVID in this
cohort successfully excluded males with Btk mutations.