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Characterization of an Uncinocarpus reesii-expressed recombinant tube precipitin antigen of Coccidioides posadasii for serodiagnosis.
PLoS One 2019; 14(8):e0221228Plos

Abstract

Early and accurate diagnosis of coccidioidomycosis, also known as Valley fever, is critical for appropriate disease treatment and management. Current serodiagnosis is based on the detection of patient serum antibodies that react with tube precipitin (TP) and complement fixation (CF) antigens of Coccidioides. IgM is the first class of antibodies produced by hosts in response to coccidioidal insults. The highly glycosylated β-glucosidase 2 (BGL2) is a major active component of the TP antigen that stimulates IgM antibody responses during early Coccidioides infection. The predominant IgM epitope on BGL2 is a unique 3-O-methyl-mannose moiety that is not produced by commonly used protein expression systems. We genetically engineered and expressed a recombinant BGL2 (rBGL2ur), derived from Coccidioides, in non-pathogenic Uncinocarpus reesii, a fungus phylogenetically related to the Coccidioides pathogen. The rBGL2ur protein was purified from the culture medium of transformed U. reesii by nickel affinity chromatography, and the presence of 3-O-methyl mannose was demonstrated by gas chromatography. Seroreactivity of the purified rBGL2ur protein was tested by enzyme-linked immunosorbent assays using sera from 90 patients with coccidioidomycosis and 134 control individuals. The sensitivity and specificity of the assay with rBGL2ur were 78.8% and 87.3%, respectively. These results were comparable to those obtained using a proprietary MiraVista Diagnostic (MVD) IgM (63.3% sensitivity; 96.3% specificity), but significantly better than the ID-TP assay using non-concentrated patient sera (33.3% sensitivity; 100% specificity). Expression of rBGL2ur in U. reesii retains its antigenicity for coccidioidomycosis serodiagnosis and greatly reduces biosafety concerns for antigen production, as Coccidioides spp. are biological safety level 3 agents.

Authors+Show Affiliations

South Texas Center for Emerging Infectious Disease and Department of Biology, University of Texas at San Antonio, San Antonio, Texas, United States of America.MiraVista Diagnostics, Indianapolis, Indiana, United States of America.South Texas Center for Emerging Infectious Disease and Department of Biology, University of Texas at San Antonio, San Antonio, Texas, United States of America.Department of Medicine, School of Medicine & Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.Department of Medicine, School of Medicine & Public Health, University of Wisconsin-Madison, Madison, Wisconsin, United States of America.MiraVista Diagnostics, Indianapolis, Indiana, United States of America.Department of Medicine, University of Arizona College of Medicine, Tucson, Arizona, United States of America.South Texas Center for Emerging Infectious Disease and Department of Biology, University of Texas at San Antonio, San Antonio, Texas, United States of America.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31412087

Citation

Yu, Jieh-Juen, et al. "Characterization of an Uncinocarpus Reesii-expressed Recombinant Tube Precipitin Antigen of Coccidioides Posadasii for Serodiagnosis." PloS One, vol. 14, no. 8, 2019, pp. e0221228.
Yu JJ, Holbrook E, Liao YR, et al. Characterization of an Uncinocarpus reesii-expressed recombinant tube precipitin antigen of Coccidioides posadasii for serodiagnosis. PLoS ONE. 2019;14(8):e0221228.
Yu, J. J., Holbrook, E., Liao, Y. R., Zarnowski, R., Andes, D. R., Wheat, L. J., ... Hung, C. Y. (2019). Characterization of an Uncinocarpus reesii-expressed recombinant tube precipitin antigen of Coccidioides posadasii for serodiagnosis. PloS One, 14(8), pp. e0221228. doi:10.1371/journal.pone.0221228.
Yu JJ, et al. Characterization of an Uncinocarpus Reesii-expressed Recombinant Tube Precipitin Antigen of Coccidioides Posadasii for Serodiagnosis. PLoS ONE. 2019;14(8):e0221228. PubMed PMID: 31412087.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of an Uncinocarpus reesii-expressed recombinant tube precipitin antigen of Coccidioides posadasii for serodiagnosis. AU - Yu,Jieh-Juen, AU - Holbrook,Eric, AU - Liao,Yu-Rou, AU - Zarnowski,Robert, AU - Andes,David R, AU - Wheat,L Joseph, AU - Malo,Joshua, AU - Hung,Chiung-Yu, Y1 - 2019/08/14/ PY - 2019/04/03/received PY - 2019/08/01/accepted PY - 2019/8/15/entrez PY - 2019/8/15/pubmed PY - 2019/8/15/medline SP - e0221228 EP - e0221228 JF - PloS one JO - PLoS ONE VL - 14 IS - 8 N2 - Early and accurate diagnosis of coccidioidomycosis, also known as Valley fever, is critical for appropriate disease treatment and management. Current serodiagnosis is based on the detection of patient serum antibodies that react with tube precipitin (TP) and complement fixation (CF) antigens of Coccidioides. IgM is the first class of antibodies produced by hosts in response to coccidioidal insults. The highly glycosylated β-glucosidase 2 (BGL2) is a major active component of the TP antigen that stimulates IgM antibody responses during early Coccidioides infection. The predominant IgM epitope on BGL2 is a unique 3-O-methyl-mannose moiety that is not produced by commonly used protein expression systems. We genetically engineered and expressed a recombinant BGL2 (rBGL2ur), derived from Coccidioides, in non-pathogenic Uncinocarpus reesii, a fungus phylogenetically related to the Coccidioides pathogen. The rBGL2ur protein was purified from the culture medium of transformed U. reesii by nickel affinity chromatography, and the presence of 3-O-methyl mannose was demonstrated by gas chromatography. Seroreactivity of the purified rBGL2ur protein was tested by enzyme-linked immunosorbent assays using sera from 90 patients with coccidioidomycosis and 134 control individuals. The sensitivity and specificity of the assay with rBGL2ur were 78.8% and 87.3%, respectively. These results were comparable to those obtained using a proprietary MiraVista Diagnostic (MVD) IgM (63.3% sensitivity; 96.3% specificity), but significantly better than the ID-TP assay using non-concentrated patient sera (33.3% sensitivity; 100% specificity). Expression of rBGL2ur in U. reesii retains its antigenicity for coccidioidomycosis serodiagnosis and greatly reduces biosafety concerns for antigen production, as Coccidioides spp. are biological safety level 3 agents. SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/31412087/Characterization_of_an_Uncinocarpus_reesii-expressed_recombinant_tube_precipitin_antigen_of_Coccidioides_posadasii_for_serodiagnosis L2 - http://dx.plos.org/10.1371/journal.pone.0221228 DB - PRIME DP - Unbound Medicine ER -