PMID- 9801343
OWN - NLM
STAT- MEDLINE
DCOM- 19981223
LR  - 20210526
IS  - 1071-412X (Print)
IS  - 1098-6588 (Electronic)
IS  - 1071-412X (Linking)
VI  - 5
IP  - 6
DP  - 1998 Nov
TI  - Use of the 27-kilodalton recombinant protein from Paracoccidioides brasiliensis 
      in serodiagnosis of paracoccidioidomycosis.
PG  - 826-30
AB  - Paracoccidioidomycosis (PCM) is one of the most important endemic mycoses in 
      Latin America; it is usually diagnosed by observation and/or isolation of the 
      etiologic agent, Paracoccidioides brasiliensis, as well as by a variety of 
      immunological methods. Although the latter are effective, two circumstances, 
      cross-reactions with other mycotic agents and antigen preparation that is marked 
      by extreme variability among lots, hinder proper standardization of the 
      procedures. To circumvent this lack of reproducibility, molecular biology tools 
      were used to produce a recombinant 27-kDa-molecular-mass antigen from this 
      fungus; a sizable quantity of this antigen was obtained through fermentation of 
      Escherichia coli DH5alpha, which is capable of expressing the fungal protein. The 
      latter was purified by the Prep-Cell System (Bio-Rad); the recovery rate of the 
      pure protein was approximately 6%. A battery of 160 human serum samples, 
      consisting of 64 specimens taken at the time of diagnosis from patients with PCM 
      representing the various clinical forms plus 15 serum specimens each from 
      patients with histoplasmosis and aspergillosis, 10 each from patients with 
      cryptococcosis and tuberculosis, 6 from patients with coccidioidomycosis, and 40 
      from healthy subjects, were all tested by an indirect enzyme-linked immunosorbent 
      assay with the purified 27-kDa recombinant protein. The latter was used at a 
      concentration of 1.0 microgram/well; there were three serum dilutions (1:1,000, 
      1:2,000, and 1:4,000). The experiment was repeated at least twice. The average 
      sensitivity for both experiments was 73.4%; in comparison with the healthy 
      subjects, the specificity for PCM patients was 87.5% while for patients with 
      other mycoses, it was 58.7%. Important cross-reactions with sera from patients 
      with aspergillosis and histoplasmosis were detected. The positive predictive 
      value of the test was 90.4%. These results indicate that it is possible to employ 
      recombinant antigenic proteins for the immunologic diagnosis of PCM and, by so 
      doing, achieve high coverage rates. Furthermore, antigen reproducibility can now 
      be ensured, thus facilitating inter- and intralaboratory standardization.
FAU - Ortiz, B L
AU  - Ortiz BL
AD  - Biochemistry and Molecular Biology Units and Mycology Group, Corporacion para 
      Investigaciones Biologicas (CIB), Medellin, Colombia.
FAU - Diez, S
AU  - Diez S
FAU - Uran, M E
AU  - Uran ME
FAU - Rivas, J M
AU  - Rivas JM
FAU - Romero, M
AU  - Romero M
FAU - Caicedo, V
AU  - Caicedo V
FAU - Restrepo, A
AU  - Restrepo A
FAU - McEwen, J G
AU  - McEwen JG
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Clin Diagn Lab Immunol
JT  - Clinical and diagnostic laboratory immunology
JID - 9421292
RN  - 0 (Antibodies, Fungal)
RN  - 0 (Antigens, Fungal)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Antibodies, Fungal/*blood
MH  - Antigens, Fungal/genetics/*immunology
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Escherichia coli/genetics/growth & development/metabolism
MH  - Fermentation
MH  - Humans
MH  - Paracoccidioides/genetics/*immunology
MH  - Paracoccidioidomycosis/*diagnosis
MH  - Predictive Value of Tests
MH  - Recombinant Proteins/genetics/immunology
MH  - Sensitivity and Specificity
MH  - Transformation, Genetic
PMC - PMC96210
EDAT- 1998/11/05 00:00
MHDA- 1998/11/05 00:01
PMCR- 1998/11/01
CRDT- 1998/11/05 00:00
PHST- 1998/11/05 00:00 [pubmed]
PHST- 1998/11/05 00:01 [medline]
PHST- 1998/11/05 00:00 [entrez]
PHST- 1998/11/01 00:00 [pmc-release]
AID - 0127 [pii]
AID - 10.1128/CDLI.5.6.826-830.1998 [doi]
PST - ppublish
SO  - Clin Diagn Lab Immunol. 1998 Nov;5(6):826-30. doi: 10.1128/CDLI.5.6.826-830.1998.