Indirect enzyme-linked immunosorbent assay for detection of immunoglobulin G reactive with a recombinant protein expressed from the gene encoding the 116-kilodalton protein of Mycoplasma pneumoniae

J Clin Microbiol. 1999 Apr;37(4):1024-9. doi: 10.1128/JCM.37.4.1024-1029.1999.

Abstract

Serology remains the method of choice for laboratory diagnosis of Mycoplasma pneumoniae infection. Currently available serological tests employ complex cellular fractions of M. pneumoniae as antigen. To improve the specificity of M. pneumoniae diagnosis, a recombinant protein was assessed as a serodiagnostic reagent. A panel of recombinant proteins were expressed from a cloned M. pneumoniae gene that encodes a 116-kDa surface protein antigen. The recombinant proteins were assessed for reactivity with patient sera and the most antigenic was further assessed for its serodiagnostic potential by indirect enzyme-linked immunosorbent assay (ELISA). The ELISA based on the recombinant protein was equivalent in sensitivity to the commercial test (Serodia Myco II; Fujirebio Inc.) to which it was compared. Southern and Western blotting data suggested that the recombinant protein derived from the 116-kDa protein of M. pneumoniae could provide a species-specific diagnostic tool, although further assessment is required.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Antibody Specificity
  • Antigens, Bacterial / chemistry
  • Antigens, Bacterial / genetics
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology*
  • Base Sequence
  • Blotting, Western
  • DNA Primers / genetics
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Evaluation Studies as Topic
  • Genes, Bacterial
  • Humans
  • Immunodominant Epitopes / chemistry
  • Immunodominant Epitopes / genetics
  • Immunoglobulin G / blood*
  • Molecular Weight
  • Mycoplasma pneumoniae / genetics
  • Mycoplasma pneumoniae / immunology*
  • Pneumonia, Mycoplasma / diagnosis
  • Pneumonia, Mycoplasma / immunology
  • Polymerase Chain Reaction
  • Rabbits
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Serologic Tests
  • Species Specificity

Substances

  • Antibodies, Bacterial
  • Antigens, Bacterial
  • Bacterial Proteins
  • DNA Primers
  • Immunodominant Epitopes
  • Immunoglobulin G
  • Recombinant Fusion Proteins