Detection of 41-kDa bacterial flagellin protein by the lymphocyte transformation test-memory lymphocyte immunostimulation assay

Abstract

Background/objectives: Diagnosis of human infection by various species of the bacterial genus Borrelia is mainly reliant on serological testing, polymerase chain reaction (PCR) or culture but such serological tests have been reported to have heterogeneous sensitivities, while Borrelia PCR and culture have been reported as being of modest diagnostic value. It has been suggested that the adjunctive use of the lymphocyte transformation test-memory lymphocyte immunostimulation assay (LTT-MELISA) may be helpful in this regard; however, the clinical usefulness of this assay has been questioned. The Borrelia immunodominant 41-kDa flagellin protein almost always gives rise to a marked human antibody response following infection. It was therefore decided to determine whether the LTT-MELISA detects the human antibody response to this antigen.

Methods: Blood samples from consecutive patients with possible borreliosis attending a clinic were independently tested by both Western blots and LTT-MELISA.

Results: After omitting cases with indeterminate Western blot results and equivocal LTT-MELISA results, multiple linear regression modelling demonstrated that the 41-kDa flagellin immunoglobulin (Ig) M level was predictable from two LTT-MELISA variables (F _2,51 = 5.981, P = 0.005). Similarly, the corresponding 41-kDa IgG model also contained two LTT-MELISA variables (F _2,57 = 3.700, P = 0.031).

Conclusion: It is concluded that the LTT-MELISA appears to be able to detect the response to this antigen.

Keywords: Flagellin; MELISA; lymphocyte transformation test.