Improved sensitivity and specificity of enzyme immunoassays with P1-adhesin enriched antigen to detect acute Mycoplasma pneumoniae infection

• Published in: Journal of microbiological methods 2001-02, Vol.44 (1), p.27-37
• Main Authors: Tuuminen, Tamara, Suni, Jukka, Kleemola, Marjaana, Jacobs, Enno
• Format: Article
• Language: English
• Subjects: Acute Disease; Adhesins, Bacterial; Adolescent; Antigens, Bacterial; Bacterial diseases; Biological and medical sciences; Blotting, Western; Child; Child, Preschool; Enzyme immunoassay; Experimental bacterial diseases and models; Humans; IgA; IgG; IgM antibodies; Immunoenzyme Techniques - methods; Immunoglobulin A - blood; Immunoglobulin G - blood; Immunoglobulin M - blood; Infant; Infectious diseases; Medical sciences; Mycoplasma pneumoniae; P1 adhesin; P1-enriched antigen; Pneumonia, Mycoplasma - diagnosis; Reagent Kits, Diagnostic; Sensitivity and Specificity; Serologic Tests - methods
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/S0167-7012(00)00235-9

An in-house P1-enriched (168-kDA protein) Mycoplasma pneumoniae antigen preparation was compared in IgG, IgA and IgM enzyme immunoassays (EIAs) to the respective EIAs employing crude antigen lysate, antigen prepared by Triton X-114 partition and two commercial antigens, one of which was an ether-extracted antigen and the other a P1-enriched antigen. In addition, three commercial kits from Sanofi Pasteur, Novum Diagnostica and Savyon Diagnostics were also assessed for comparison. Diagnostic sensitivity was studied with paired samples from adults ( n=37) with acute respiratory illness interpreted as acute, recent or past infection to M. pneumoniae on the basis of the results of complement fixation test (CFT). If the consensus of at least two methods is taken as the true positive for acute infection, the diagnostic sensitivities of combined IgG and IgM EIAs were 100% for the Platelia®, Sero™ MP and in-house EIAs whereas for the Novum EIAs and CFT- 97% and 74%, respectively. Moreover, the sensitivity of the P1-enriched antigen was proven superior on the basis of systematically highest OD 405 nm ratios between convalescent and acute serum samples. Analytical specificity was studied by screening serum samples from 92 Finnish blood donors and 111 serum samples from cord blood. Diagnostic specificity was studied in a blind testing of 30 paired serum samples from infants with pneumonia of variable etiology. No single misinterpretation of acute infection from the group of samples with other respiratory diseases did occur. The present study confirmed and extended the earlier observations of the usefulness of P1-enriched antigen for reliable serologic diagnosis of acute M. pneumoniae infection.