title: High molecular mass fraction in clinical isolates of Paracoccidioides brasiliensis.
abstract: BACKGROUND
  Different serum levels of the IgG/IgE for Paracoccidioides brasiliensis high mass molecular (hMM) fraction (~366 kDa) in the acute and chronic forms of the disease have been reported. Considering the nonexistence of hMM fraction investigation involving clinical isolates of P. brasiliensis, the present study aimed to investigate the presence of the hMM fraction (~366 kDa) in cell free antigens (CFA) from P. brasiliensis clinical isolates.
  
  
  METHODS
  CFA from 10 clinical isolates and a reference strain (Pb18) were submitted to SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by gel image capturing and densitometer analysis. Additionally, CFA from 20 isolates and Pb18 were analyzed by capture ELISA (cELISA) using polyclonal (polAb) or monoclonal (mAb) antibodies to the hMM fraction.
  
  
  RESULTS
  The presence of the hMM component was observed in CFA of all samples analyzed by SDS-PAGE/densitometry and by cELISA. In addition, Pearson's correlation test demonstrated stronger coefficients between hMM fraction levels using pAb and mAb (R = 0.853) in cELISA.
  
  
  CONCLUSIONS
  The soluble hMM fraction was present in all the P. brasiliensis clinical isolates analyzed and the reference strain Pb18, which could be used as a source of this antigen. The work also introduces for first time, the cELISA method for P. brasiliensis hMM fraction detection. Analysis also suggests that detection is viable using polAb or mAb and this methodology may be useful for future investigation of the soluble hMM fraction (~366 kDa) in sera from PCM patients.
authors.affiliation: Department of Pathology Science, Biological Science Center, State University of Londrina, Londrina, PR, Brazil.
authors.forename: Andréa Longoni
authors.initials: AL
authors.lastname: Fredrich
authors.forename: Luciene Airy
authors.initials: LA
authors.lastname: Nagashima
authors.forename: Wander Rogério
authors.initials: WR
authors.lastname: Pavanelli
authors.forename: Audrey de Souza
authors.initials: Ade S
authors.lastname: Marquez
authors.forename: Mari Sumigawa
authors.initials: MS
authors.lastname: Kaminami
authors.forename: Nilson de Jesus
authors.initials: Nde J
authors.lastname: Carlos
authors.forename: Ayako
authors.initials: A
authors.lastname: Sano
authors.forename: Mario Augusto
authors.initials: MA
authors.lastname: Ono
authors.forename: Eiko Nakagawa
authors.initials: EN
authors.lastname: Itano
mesh_terms: D000908:Antibodies, Fungal / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000946:Antigens, Fungal / Q000276:immunology*; D002908:Chronic Disease; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D008970:Molecular Weight; D010228:Paracoccidioides / Q000276:immunology* / Q000302:isolation & purification; D010229:Paracoccidioidomycosis / Q000469:parasitology*
chemical_list: D000908:Antibodies, Fungal; D000911:Antibodies, Monoclonal; D000946:Antigens, Fungal; D007074:Immunoglobulin G