Direct Detection of Mycoplasma pneumoniae Antigen in Clinical Specimens by a Monoclonal Antibody Immunoblot Assay

Abstract

Throat swabs from patients with pharyngitis and sputum specimens from patients with atypical pneumonia were tested for the presence of a Mycoplasma pneumoniae polypeptide with a molecular weight of 43,000 with the use of an M. pneumoniae species-specific monoclonal antibody in an immunoblot assay. This 43,000-dalton polypeptide was detectable in 33 of 33 throat swabs from patients with pharyngitis that were positive for M. pneumoniae by conventional culture as well as a culture-amplified enzyme immunoassay. The 43,000-dalton polypeptide was also detected in three of three M. pneumoniae culture-positive sputum specimens. It was not detected in 3 sputum specimens culture-confirmed for Legionella pneumophila, 10 sputum specimens from normal persons, or 25 throat swabs also from normal persons. This immunoblot assay could be completed within five hours and may be an alternative method for detecting M. pneumoniae antigen directly in sputum or throat swab specimens.

A Latex Agglutination Test for the Detection of Mycoplasma pneumoniae in Respiratory Exudates: A Comparative Study with a Commercially Available DNA‐Probe Test

“…In this paper, we described a latex agglutination test for detecting directly M. pneumoniae antigens in respiratory exudates. For the direct detection and rapid diagnosis of M. pneumoniae, the DNA-probe method (DP) (6, 8-11, 16, 17, 22, 27), the polymerase chain reaction (PCR) (3), the monoclonal antibody immunoblot assay (21), the antigen capture enzyme immunoassay (Ag-EIA) (10,18,30) and the indirect immunofluorence (13,14) have recently been developed. Among these methods, only DP is commercially available, and the kit of DP has been evaluated with clinical specimens by many investigators.…”

Diagnosis of Mycoplasma pneumoniae pneumonia: sensitivities and specificities of serology with lipid antigen and isolation of the organism on soy peptone medium for identification of infections

“…The detection limit of the polyclonal rabbit antiserum assay of Kok et al (31) was 104 CFU from culture. Two different monoclonal antibodies against a 43-kilodalton antigen (7,32) have been used for antigen detection. One test had a detection limit of 105 to 106 CFU, and thus it was necessary to culture the specimens in medium for 2 to 12 days to amplify the amount of antigen sufficiently for detection (7).…”

Chlamydia pneumoniaeandMycoplasma pneumoniae

“…Monoclonal antibody immunoblot A has been used in research samples for M. pneumoniae antigen detection in respiratory specimens. 25 Commercially available DNA and RNA probes show high specificity but low sensitivity (22-100%) in pharyngeal specimens. 26 The polymerase chain reaction (PCR) has been applied to clinical specimens showing the suitability of this technique for the detection of M. pneumoniae.…”