IDENTIFICATION OFMYCOPLASMATACEAEBY THE FLUORESCENT ANTIBODY METHOD

Abstract

BROWN. Identification of Mllycoplasmataceae by the fluorescent antibody method. J. Bacteriol. 85:111-118. 1963.-The conditions of the fluorescent antibody reactions were studied in relation to their application to Mycoplasmataceae or pleuropneumonia-like organisms (PPLO). Aliycoplasma hominis type 1 and 2 antigens and their homologous antisera were used to determine the activity and specificity of these and other strains. Fluorescein isothiocyanate conjugated antiserum globulin preparations were used in both the direct and indirect fluorescent antibody methods. A direct tube technique was used for the detection and measurement of growth in broth cultures by the addition of conjugated antiserum. The specific fluorescent staining and recognition of hot water fixed M. hominis colonies was presented as a suitable identification standard. The antigenic activity was found to remain in the insoluble residue after exposure of 31. hominis strains to sonic vibration (9 ke) for 30 min and centrifugation. Brief 2-min exposures of tissue cells to vibration (9 ke) caused the disruption of tissues, with the release of viable and "bound" nonwashable strains that reacted

GROWTH INHIBITORY PROPERTIES OF MYCOPLASMA ANTIBODY

“…Recently Clark et al (1963) demonstrated that fluorescent antibody remained fixed to the MIycoplasma membrane residue after exposure to sonic oscillation. Our additional observation that viable Mlycoplasma can be recovered from antibody complex by sonic oscillation appears to exclude lysis as the primary cause of growth inhibition.…”

“…Our recovery of viable MIycoplasma from apparently negative antiserum broth cultures after exposure to sonic oscillation suggests a latent phase of the organisms in a neutralized form. Clark et al (1963) also used sonic oscillation to release viable Mlycoplasma "bound" in tissue cell cultures. In view of these findings, further investigation of the application of sonic oscillation techniques to bodY fluids and tissues suspected of containing neutralized JMycoplasma appears to be indicated.…”

Starch Gel Electrophoresis of a Fraction of Certain of the Pleuropneumonia-Like Group of Microorganisms

“…Various strains of the PPLO, isolated in several laboratories (Table 1) and maintained in a fluid medium described previously (Brown, 1948;Clark et al, 1963), were used. The medium consisted of a pancreatic digest of beef heart enriched with 10% heat-inactivated (56 C for 30 FOWLER ET AL.…”

“…Antigenic variations among the pleuropneumonia-like group of microorganisms (PPLO), defined herein as including various "L" forms of bacteria and the Mycoplasmataceae, have been repeatedly demonstrated by various serological reactions (Bailey et al, 1961;Card, 1959;Clark et al, 1963;Coriell, Fabrizio, and Wilson, 1960;Klieneberger, 1938Klieneberger, , 1940Malizia, Barile, and Riggs, 1961). This, together with rather striking host specificity and often tissue localization, suggests that some physicochemically demonstrable differences may exist in the proteins of the different species.…”

“…The successful cultivation of large quantities of the microorganisms by use of continuous agitation (Clark et al, 1963) has made possible the utilization of the starch gel technique (Pert et al, 1959;Poulik, 1957;Smithies, 1959) for the further characterization of the electrophoretically mobile protein components of the PPLO.…”