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Updated: Aug 9, 2026
Antigen-Capture Enzyme-Linked Immunosorbent Assay for Specific Detection of Mycoplasma pneumoniae
Published on: February 24, 2023
Isolation, Characterization, and Immunogenicity of Mycoplasma pneumoniae Membranes
J D Pollack1, N L Somerson, L B Senterfit
1Departments of Medical Microbiology and Pediatrics, Ohio State University, Children's Hospital, Columbus, Ohio 43205.
Abstract:
Membrane and soluble fractions of Mycoplasma pneumoniae, M. pulmonis, and M. laidlawii B were prepared by hypotonic lysis of whole cells. The membranes of M. pneumoniae and M. laidlawii B contained, as percentage of dry weight: 34 to 37% protein, 59 to 61% lipid, 3 to 4% carbohydrate as hexose, and 0.2% ribonucleic acid as ribose. NADH(2) and NADPH(2) oxidase activities were localized in the soluble fractions of M. pneumoniae and in the membrane fraction of M. laidlawii B. NADH(2) oxidase activity was localized in the soluble fraction of M. pulmonis. The lipids of M. pneumoniae were labeled when the organism was grown in the presence of either radioactive palmitic acid, oleic acid, cholesterol, or glycerol. The lipids were not labeled when grown in the presence of radioactive acetate. Palmitic acid radio-activity was found in neutral lipid, glycolipid, and phosphatide fractions. Immunodiffusion analyses of whole cells and membrane fractions demonstrated three reactive antigens. Two immunodiffusion antigens were localized in the membrane fraction. One of these apparently contains lipid. A third antigen, also considered lipoidal, was found in whole cells. Membrane and soluble fractions of M. pneumoniae were immunogenic. The immunogens eliciting metabolic-inhibiting antibodies were localized in the membrane. The membrane preparation also induced the formation of antibodies which fixed complement with an antigen extracted with lipid solvent. The soluble fraction contained a distinct immunogen which induces antibodies reactive in complement fixation with an antigen prepared by phenol extraction.
Insights
This study characterizes the membrane and soluble fractions of Mycoplasma species, identifying key components and their locations. It reveals specific immunogens within Mycoplasma pneumoniae membranes responsible for metabolic-inhibiting and complement-fixing antibodies.
Area of Science:
- Microbiology
- Cell Biology
- Immunology
Background:
- Mycoplasma species are cell-wall-less bacteria with unique membrane compositions.
- Understanding their cellular fractions is crucial for comprehending their biology and pathogenicity.
Purpose of the Study:
- To analyze the biochemical composition and enzymatic activities of membrane and soluble fractions from Mycoplasma pneumoniae, M. pulmonis, and M. laidlawii B.
- To identify and characterize immunogenic components within these fractions, particularly those of Mycoplasma pneumoniae.
Main Methods:
- Hypotonic lysis was used to prepare membrane and soluble cellular fractions.
- Biochemical analysis determined protein, lipid, carbohydrate, and RNA content.
- Enzymatic activities (NADH(2) and NADPH(2) oxidase) were localized.
- Lipid labeling studies incorporated radioactive precursors.
- Immunodiffusion and complement fixation assays identified antigens and immunogens.
Main Results:
- Mycoplasma pneumoniae and M. laidlawii B membranes are rich in lipids and proteins.
- NADH(2) and NADPH(2) oxidase activities were found in different cellular fractions depending on the species.
- Lipids of M. pneumoniae incorporated various fatty acids and sterols, with palmitic acid found in neutral lipid, glycolipid, and phosphatide fractions.
- Three reactive antigens were detected, with two localized in the membrane, one possibly lipid-containing.
- Both membrane and soluble fractions of M. pneumoniae were immunogenic, with membrane components eliciting metabolic-inhibiting and complement-fixing antibodies.
Conclusions:
- The study delineates the distinct biochemical and antigenic profiles of Mycoplasma cellular fractions.
- Membrane-associated components of Mycoplasma pneumoniae are significant immunogens, capable of inducing antibodies that interfere with cellular metabolism and activate complement.
- These findings contribute to understanding Mycoplasma-host interactions and potential vaccine development.
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