Immunofluorescence

Abstract

Given the application of IFA techniques to immunodiagnostics in other areas, it was an anticipated extrapolation that similar methods should be applied to mycoplasmology. Essentially, the IFA approach provided the perceived opportunity of enhancing sensitivity and specificity at a time when competing methods were limited. The availability of suitable fluorescein-conjugated antibodies was also a limiting factor for routine IFA serology but standardization in this area quickly overcame this dilemma. The critical experiments of Liu et al.⁹ had proposed an association of primary atypical pneumonia and the “Mac virus” through the application of IFA serology which was based on antibody recognition of antigen from infected chick embryo lung sections. Shortly thereafter, the IFA technique using infected chick tissue sections was of value in furthering the science⁷ of primary atypical pneumonia causation. In the early stages, the derivation and use of other serological techniques, especially CF, was dependent on the availability of IFA as a reference.^120–122 In fact, this specific IFA procedure continued to be utilized for almost a decade thereafter.^27,123,124 Although the concept of using immunofluorescence continued to appear feasible, the antigenic substrate, i.e. tissues sections, was cumbersome and relatively less transferable.