PMID- 21354162
OWN - NLM
STAT- MEDLINE
DCOM- 20110602
LR  - 20220311
IS  - 1872-7905 (Electronic)
IS  - 0022-1759 (Linking)
VI  - 367
IP  - 1-2
DP  - 2011 Mar 31
TI  - Combined use of Paracoccidioides brasiliensis recombinant rPb27 and rPb40 
      antigens in an enzyme-linked immunosorbent assay for immunodiagnosis of 
      paracoccidioidomycosis.
PG  - 78-84
LID - 10.1016/j.jim.2011.02.006 [doi]
AB  - Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in 
      Latin America; it's usually diagnosed by observation and/or isolation of the 
      etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of 
      immunological methods, such as complement fixation and immunodiffusion. Although 
      these approaches are useful, historically their sensitivity and specificity have 
      often been compromised by the use of complex mixtures of undefined antigens. The 
      use of combinations of purified, well-characterized antigens appears preferable 
      and may yield good results. In the present study combinations of the previously 
      described 27-kDa recombinant antigen (rPb27) and a recombinant 
      40-kDa-molecular-mass antigen (rPb40) from this fungus, that was identified by 
      Goes et al. (2005) through the AST strategy as a homolog of Neurospora crassa 
      calcineurin B, were used in an indirect enzyme linked immunosorbent assay (ELISA) 
      for diagnosis and follow-up of patients with PCM. The complete coding cDNA of 
      rPb40 and rPb27 were cloned into a pET-21a and a pET-DEST 42 plasmid, 
      respectively, expressed in E. coli with a his-tag and purified by affinity 
      chromatography. Among 109 PCM serum samples analyzed, a homogeneous IgG response 
      to these proteins was observed. 62 serum samples from patients with other 
      diseases, 18 from patients with other mycosis and 23 from healthy individuals 
      were also studied. Detection of anti-rPb27 and anti-rPb40 antibodies in sera of 
      patients with PCM by ELISA using a combination of the two purified proteins 
      showed a sensitivity of 96% with a specificity of 100% in relation to control 
      normal human sera and to sera from patients with other systemic mycosis and 93.5% 
      to sera from patients with diverse infections. The use of this two proteins 
      combination provided an excellent immunodiagnosis assay with great values of 
      sensitivity and specificity, even in relation to sera from patients with other 
      mycosis, making possible the standardization of a new methodology to diagnose 
      this important mycosis, with a good confiability and reprodutibility.
CI  - Copyright (c) 2011 Elsevier B.V. All rights reserved.
FAU - Fernandes, V C
AU  - Fernandes VC
AD  - Departamento de Bioquimica e Imunologia, Instituto de Ciencias Biologicas, UFMG, 
      Belo Horizonte, MG, Brazil.
FAU - Coitinho, J B
AU  - Coitinho JB
FAU - Veloso, J M R
AU  - Veloso JM
FAU - Araujo, S A
AU  - Araujo SA
FAU - Pedroso, E P
AU  - Pedroso EP
FAU - Goes, A M
AU  - Goes AM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110224
PL  - Netherlands
TA  - J Immunol Methods
JT  - Journal of immunological methods
JID - 1305440
RN  - 0 (Antibodies, Fungal)
RN  - 0 (Antigens, Fungal)
RN  - 0 (Immunoglobulin G)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Aged
MH  - Antibodies, Fungal/*blood
MH  - Antigens, Fungal/*immunology
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Humans
MH  - Immunoglobulin G/blood
MH  - Middle Aged
MH  - Paracoccidioides/*immunology
MH  - Paracoccidioidomycosis/*diagnosis/immunology
MH  - Recombinant Proteins/immunology
MH  - Sensitivity and Specificity
EDAT- 2011/03/01 06:00
MHDA- 2011/06/03 06:00
CRDT- 2011/03/01 06:00
PHST- 2010/12/28 00:00 [received]
PHST- 2011/02/15 00:00 [revised]
PHST- 2011/02/17 00:00 [accepted]
PHST- 2011/03/01 06:00 [entrez]
PHST- 2011/03/01 06:00 [pubmed]
PHST- 2011/06/03 06:00 [medline]
AID - S0022-1759(11)00038-X [pii]
AID - 10.1016/j.jim.2011.02.006 [doi]
PST - ppublish
SO  - J Immunol Methods. 2011 Mar 31;367(1-2):78-84. doi: 10.1016/j.jim.2011.02.006. 
      Epub 2011 Feb 24.