[Development of serodiagnostic kit "HITAZYME Chlamydia Ab" for Chlamydia trachomatis infections using extracted antigen].
To develop EIA kit with low cross-reactivity for the quantitative detection of anti-chlamydial antibodies, we examined the preparation of trachomatis antigens and its specificity to mouse antisera and human sera. The chlamydial elementary body (EB) purified from C. trachomatis L2/434/Bu strain was treated by Sarkosyl, dithiothreitol and SDS by turns to obtain the soluble EB outer membrane (COMC). SDS-PAGE showed that the major components of the COMC were 96K, 60K and 39.5 KDa peptides. The reactivity of the COMC immobilized to 96 wells microtiter plate to mouse anti-serum to C. trachomatis was higher than the other two mouse anti-sera to C. psittaci and pneumoniae. In human sera, the cut off values were calculated from an average optical density plus its two-fold standard deviation obtained by the testing of 100 samples of healthy human sera. We evaluated the specificity of the kit to 17 anti-C. pneumoniae, 9 C. trachomatis and 4 C. psittaci antibodies positive patients' sera judged by the MFA method respectively. The results showed that the concordance ratio of IgG and IgA were 88%, 100% in anti-C. pneumoniae, 89%, 78% in anti-C. trachomatis and 50%, 50% in anti-C. psittaci respectively. From the results obtained in this study, we concluded that the HITAZYME method which had a very low cross-reactivity to C. pneumoniae is clinically useful in the serodiagnosis of C. trachomatis infections, even if it has a little common antigenicity with C. psittaci antigen.
Matsumoto
A
A
Department of Microbiology, Kawasaki Medical School, Kurashiki, Japan.
Bessho
H
H
Kishimoto
T
T
Soejima
R
R
Watanabe
H
H
Kawagoe
K
K
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis*; D000942:Antigens, Bacterial / Q000276:immunology*; D002648:Child; D002690:Chlamydia Infections / Q000175:diagnosis* / Q000276:immunology; D002692:Chlamydia trachomatis / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D008297:Male; D051379:Mice; D008875:Middle Aged; D011933:Reagent Kits, Diagnostic
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D000939:Epitopes; D011933:Reagent Kits, Diagnostic