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Appl Environ Microbiol. 1974 February; 27(2): 356-359
Copyright © 1974 American Society for Microbiology. All Rights Reserved.

Stable Mycoplasma Antigen Preparations for Indirect Hemagglutination Tests

¹ William Pepper Laboratory, Microbiology Division, Department of Pathology, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19174

ABSTRACT

In immunological studies of mycoplasmas, the use of glutaraldehyde for the fixative makes it possible to use erythrocytes from commercially available defibrinated sheep blood. It eliminates the necessity of having to screen blood from individual sheep to obtain a suitable source of erythrocytes, as when employing tannic acid for fixation and sensitization. The chemical bonding of soluble mycoplasma proteins to glutaraladehyde-fixed sheep erythrocytes by bis-diazotized 3,3'dimethoxy derivative, benzidine, yields preparations that are satisfactory antigens for performing the indirect hemagglutination test by the microtiter technique. The antigenic preparations are satisfactory for use after storage at 4 or -10 C for many months. Incorporation of 5% glycerine in the final suspending milieu makes it possible to obtain uniform suspensions of the fixed and sensitized sheep erythrocytes after freezing and after repeated freezing and thawing. Proteins from Mycoplasma arthritidis and M. hominis have been coupled to glutaraldehyde-fixed erythrocytes by diazotization. The last mentioned preparation detected the presence of antibodies in titers greater than 1:10 in 37% of 237 pregnant women whose ages ranged between 20 and 30 years. There was no correlation between the presence of specific antibodies in the blood and the isolation of M. hominis from the cervical canal.