Preparation of Pleuropneumonia-Like Organisms for Microscopic Study

Clark, Harold W.; Fowler, Richard C.; Brown, Thomas McP.

doi:10.1128/jb.81.3.500-502.1961

Cited by 35 publications

(5 citation statements)

References 0 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunofluorescence technique. The PPLO colonies were transferred from agar plates to glass slides by a technique previously described (Clark, Fowler, and Brown, 1961). Normal or immune rabbit serum (diluted in 10% guinea pig complement) was added to the slides.…”

Section: Methodsmentioning

confidence: 99%

Serological Relationships Among Human Mycoplasmas as Shown by Complement-Fixation and Gel Diffusion

et al. 1963

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Serological Relationships Among Human Mycoplasmas as Shown by Complement-Fixation and Gel Diffusion

et al. 1963

View full text Add to dashboard Cite

show abstract

“…The supernatants were examined for viruses by standard methods and for M. pneumoniae by inoculation of diphasic media with agar subculture or a HeLa 229 cell monolayer culture-agar subculture system (Kok, Varkanis & Marmion, unpublished observations). Subcultures from the HeLa cell monolayers or other media were made on Hayflick and other mycoplasma agar and suspect colonies of M. pneumoniae were identified initially by colonyhaemadsorption and then by IF staining of colonies transferred to slides by the method of Clark, Fowler & Brown (1961). Identification was also made by Ag-EIA, as detailed above.…”

Section: Culture Of Clinical Specimens For Viruses and M Pneumoniaementioning

confidence: 99%

Laboratory diagnosis ofMycoplasma pneumoniaeinfection: 1. Direct detection of antigen in respiratory exudates by enzyme immunoassay.

et al. 1988

View full text Add to dashboard Cite

Direct and indirect antigen capture enzyme immunoassays (Ag-EIA) have been developed for the detection of Mycoplasma pneumoniae in nasopharyngeal aspirates or sputum from respiratory infection. The sensitivity of the two Ag-EIA were similar, but the indirect method using polyclonal rabbit and guinea-pig antisera was more convenient. The Ag-EIA had a detection limit of 10(4-4.5) colony-forming units/ml of sample. It was specific for M. pneumoniae and gave a low level response with M. genitalium. There were no cross-reactions with 10 other species of mycoplasmas. Tests with a wide range of bacteria and chlamydia group antigen, representing agents sometimes found in the respiratory tract, were also negative. At the current level of development, the Ag-EIA detected about 90% of specimens that were also positive for culture; 43% of specimens from culture-negative--seropositive patients gave a positive result. The overall pattern of results indicated that while antigen detection is a quick and effective substitute for the slow culture method, serological examination for specific IgM antibody is also necessary to give a complete diagnostic coverage.

show abstract

“…10 min. PPLO colonies, also used in this study, were transferred and fixed directly to slides by the specially designed hot water fixation technique of Clark, Fowler, and Brown (1961). Hot distilled water fixed the proteinaceous PPLO on the slide as it dissolved away the surrounding agar, leaving the colonies relatively free of extraneous substances, and thereby reducing nonspecific fluorescence.…”

Section: Methodsmentioning

confidence: 99%

IDENTIFICATION OFMYCOPLASMATACEAEBY THE FLUORESCENT ANTIBODY METHOD

et al. 1963

Self Cite

View full text Add to dashboard Cite

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

show abstract

Preparation of Pleuropneumonia-Like Organisms for Microscopic Study

Cited by 35 publications

References 0 publications

Serological Relationships Among Human Mycoplasmas as Shown by Complement-Fixation and Gel Diffusion

Serological Relationships Among Human Mycoplasmas as Shown by Complement-Fixation and Gel Diffusion

Laboratory diagnosis ofMycoplasma pneumoniaeinfection: 1. Direct detection of antigen in respiratory exudates by enzyme immunoassay.

IDENTIFICATION OFMYCOPLASMATACEAEBY THE FLUORESCENT ANTIBODY METHOD

Contact Info

Product

Resources

About