title: [A double-antigen sandwich ELISA for detecting Penicillium marneffei Mp1p-specific antibody].
abstract: OBJECTIVE
  To establish an immunological method for detecting antibodies of Penicillium marneffei.
  
  
  METHODS
  The recombinant Mp1p protein of Penicillium marneffei was expressed in Pichia pastoris and labeled with HRP (Mp1p-HRP) with a modified sodium periodate method. A double-antigen sandwich enzyme-linked immunosorbant assay (ELISA) was established by determining the optimal coating concentration of Mp1p protein and the concentration of the detecting protein Mp1p-HRP. The sensitivity and specificity of the assay was evaluated by detecting Mp1p antibodies in 100 serum samples from healthy donors, 15 samples from culture-confirmed penicilliosis patients, and 21 samples from patients with culture-confirmed other fungal infections.
  
  
  RESULTS
  A double-antigen sandwich ELISA was successfully established for detecting Mp1p-specific antibody. The specificity of the assay was 100% (121/121) for detecting Mp1p-specific antibody in the sera from healthy donors and patients with other fungal infection. The detection results of the 15 serum samples from patients with culture-confirmed penicilliosis showed positivity for Mp1p antibody in 2 samples and Mp1p antigen positivity in 12 samples; combining the detection results of Mp1p antigen and antibody obviously increased the diagnostic sensitivity to 93.3% (14/15).
  
  
  CONCLUSIONS
  The double-antigen sandwich ELISA shows a high specificity in detecting Mp1p-specific antibody, and simultaneous detection of Mp1p antigen and antibody can increase the diagnostic sensitivity for penicilliosis.
authors.affiliation: Center for Clinical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. gzwyf2010@163.com
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Lei
authors.initials: L
authors.lastname: Zeng
authors.forename: Xuedong
authors.initials: X
authors.lastname: Chen
authors.forename: Wei
authors.initials: W
authors.lastname: Hao
authors.forename: Mei
authors.initials: M
authors.lastname: Yang
authors.forename: Jianpiao
authors.initials: J
authors.lastname: Cai
authors.forename: Yadi
authors.initials: Y
authors.lastname: Wang
authors.forename: Guoyong
authors.initials: G
authors.lastname: Yuan
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Che
mesh_terms: D000908:Antibodies, Fungal / Q000097:blood* / Q000276:immunology; D000946:Antigens, Fungal / Q000097:blood / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D009181:Mycoses / Q000097:blood* / Q000175:diagnosis / Q000382:microbiology; D010407:Penicillium / Q000276:immunology; D010843:Pichia / Q000276:immunology*; D012680:Sensitivity and Specificity
chemical_list: D000908:Antibodies, Fungal; D000946:Antigens, Fungal