Immunofluorescence has proved useful in the identification of mycoplasmas and detection of antibodies against them (1-3). For Mycoplasma pfieumoniae, the indirect fluorescent antibody test has been found to be highly specific and high fluorescent antibody titers to M . pneumoniae have been correlated with resistance to clinical illness (4). The purpose of this study was to develop antigenic preparations for a variety of mycoplasma species which would be both reliable and convenient to use in fluorescent antibody tests. Two methods of preparing mycoplasma antigens for fluorescence tests have thus far been reported. One, as described by Liu ( S ) , employs frozen sections of chick embryo lung infected with M . ifmeumoniae. In the second, and most commonly used precedure (6), mycoplasma colonies growing on agar are transferred to microscope slides and fixed in a hot water bath. The Liu technique employing egg-grown antigens is technically difficult and time consuming; the procedure utilizing hot water fixation gives nonspecific fluorescence primarily because of the presence of contaminating agar.Broth-grown mycoplasmas have been shown to adhere to glass surfaces (7). We have exploited this property by using glass cov-
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