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Article: Immunoassays based on Penicillium marneffei Mp1p derived from Pichia pastoris expression system for diagnosis of penicilliosis
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TitleImmunoassays based on Penicillium marneffei Mp1p derived from Pichia pastoris expression system for diagnosis of penicilliosis
 
AuthorsWang, YF3
Cai, JP3
Wang, YD3
Dong, H1
Hao, W3
Jiang, LX3
Long, J3
Chan, C2
Woo, PCY2
Lau, SKP2
Yuen, KY2
Che, XY3
 
Issue Date2011
 
PublisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
 
CitationPLoS One, 2011, v. 6 n. 12, article no. e28796 [How to Cite?]
DOI: http://dx.doi.org/10.1371/journal.pone.0028796
 
AbstractBACKGROUND: Penicillium marneffei is a dimorphic fungus endemic in Southeast Asia. It can cause fatal penicilliosis in humans, particularly in HIV-infected people. Diagnosis of this infection is difficult because its clinical manifestations are not distinctive. Specialized laboratory tests are necessary to establish a definitive diagnosis for successful management. We have demonstrated previously that a cell wall mannoprotein Mp1p, abundant in P. marneffei, is a potential biomarker for diagnosis of P. marneffei infections. In the present study, we describe immunoassays based on Mp1p derived from the yeast Pichia pastoris expression system. METHODOLOGY/PRINCIPAL FINDINGS: We generated monoclonal antibodies (MAbs) and rabbit polyclonal antibodies (PAbs) against Mp1p expressed in P. pastoris. Subsequently, we developed two Mp1p antigen capture ELISAs which employed MAbs for both the capture and detecting antibodies (MAb-MAb pair) or PAbs and MAbs as the capture and detecting antibodies (PAbs-MAb pair) respectively. The two Mp1p antigen ELISAs detected Mp1p specifically in cultures of P. marneffei yeast phase at 37-40 degrees C and had no cross-reaction with other tested pathogenic fungi. The sensitivities and specificities of the two antigen assays were found to be 55% (11/20) and 99.6% (538/540) for MAb-MAb Mp1p ELISA, and 75% (15/20) and 99.4% (537/540) for PAbs-MAb Mp1p ELISA performed using 20 sera with culture-confirmed penicilliosis, and 540 control sera from 15 other mycosis patients and 525 healthy donors. Meanwhile, we also developed an anti-Mp1p IgG antibody ELISA with an evaluated sensitivity of 30% (6/20) and a specificity of 98.5% (532/540) using the same sera. Furthermore, combining the results of Mp1p antigen and antibody detection improved the sensitivity of diagnosis to 100% (20/20). CONCLUSIONS/SIGNIFICANCE: Simultaneous detection of antigen and antibody using the immunoassays based on Mp1p derived from P. pastoris greatly improves detection sensitivity. The procedures should be useful for the routine diagnosis of penicilliosis.
 
ISSN1932-6203
2013 Impact Factor: 3.534
2013 SCImago Journal Rankings: 1.724
 
DOIhttp://dx.doi.org/10.1371/journal.pone.0028796
 
PubMed Central IDPMC3244411
 
ISI Accession Number IDWOS:000299113600035
Funding AgencyGrant Number
Ted Sun Foundation, Hong Kong
Research and Development Project of Guang Dong Province2010B090400498
Funding Information:

This work was supported by the Ted Sun Foundation, Hong Kong and the Research and Development Project of Guang Dong Province (No. 2010B090400498). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

 
ReferencesReferences in Scopus
 
DC FieldValue
dc.contributor.authorWang, YF
 
dc.contributor.authorCai, JP
 
dc.contributor.authorWang, YD
 
dc.contributor.authorDong, H
 
dc.contributor.authorHao, W
 
dc.contributor.authorJiang, LX
 
dc.contributor.authorLong, J
 
dc.contributor.authorChan, C
 
dc.contributor.authorWoo, PCY
 
dc.contributor.authorLau, SKP
 
dc.contributor.authorYuen, KY
 
dc.contributor.authorChe, XY
 
dc.date.accessioned2012-08-08T08:52:03Z
 
dc.date.available2012-08-08T08:52:03Z
 
dc.date.issued2011
 
dc.description.abstractBACKGROUND: Penicillium marneffei is a dimorphic fungus endemic in Southeast Asia. It can cause fatal penicilliosis in humans, particularly in HIV-infected people. Diagnosis of this infection is difficult because its clinical manifestations are not distinctive. Specialized laboratory tests are necessary to establish a definitive diagnosis for successful management. We have demonstrated previously that a cell wall mannoprotein Mp1p, abundant in P. marneffei, is a potential biomarker for diagnosis of P. marneffei infections. In the present study, we describe immunoassays based on Mp1p derived from the yeast Pichia pastoris expression system. METHODOLOGY/PRINCIPAL FINDINGS: We generated monoclonal antibodies (MAbs) and rabbit polyclonal antibodies (PAbs) against Mp1p expressed in P. pastoris. Subsequently, we developed two Mp1p antigen capture ELISAs which employed MAbs for both the capture and detecting antibodies (MAb-MAb pair) or PAbs and MAbs as the capture and detecting antibodies (PAbs-MAb pair) respectively. The two Mp1p antigen ELISAs detected Mp1p specifically in cultures of P. marneffei yeast phase at 37-40 degrees C and had no cross-reaction with other tested pathogenic fungi. The sensitivities and specificities of the two antigen assays were found to be 55% (11/20) and 99.6% (538/540) for MAb-MAb Mp1p ELISA, and 75% (15/20) and 99.4% (537/540) for PAbs-MAb Mp1p ELISA performed using 20 sera with culture-confirmed penicilliosis, and 540 control sera from 15 other mycosis patients and 525 healthy donors. Meanwhile, we also developed an anti-Mp1p IgG antibody ELISA with an evaluated sensitivity of 30% (6/20) and a specificity of 98.5% (532/540) using the same sera. Furthermore, combining the results of Mp1p antigen and antibody detection improved the sensitivity of diagnosis to 100% (20/20). CONCLUSIONS/SIGNIFICANCE: Simultaneous detection of antigen and antibody using the immunoassays based on Mp1p derived from P. pastoris greatly improves detection sensitivity. The procedures should be useful for the routine diagnosis of penicilliosis.
 
dc.description.naturepublished_or_final_version
 
dc.identifier.citationPLoS One, 2011, v. 6 n. 12, article no. e28796 [How to Cite?]
DOI: http://dx.doi.org/10.1371/journal.pone.0028796
 
dc.identifier.doihttp://dx.doi.org/10.1371/journal.pone.0028796
 
dc.identifier.hkuros204308
 
dc.identifier.isiWOS:000299113600035
Funding AgencyGrant Number
Ted Sun Foundation, Hong Kong
Research and Development Project of Guang Dong Province2010B090400498
Funding Information:

This work was supported by the Ted Sun Foundation, Hong Kong and the Research and Development Project of Guang Dong Province (No. 2010B090400498). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

 
dc.identifier.issn1932-6203
2013 Impact Factor: 3.534
2013 SCImago Journal Rankings: 1.724
 
dc.identifier.issue12, article no. e28796
 
dc.identifier.pmcidPMC3244411
 
dc.identifier.pmid22205971
 
dc.identifier.scopuseid_2-s2.0-83755186102
 
dc.identifier.urihttp://hdl.handle.net/10722/157663
 
dc.identifier.volume6
 
dc.languageeng
 
dc.publisherPublic Library of Science. The Journal's web site is located at http://www.plosone.org/home.action
 
dc.publisher.placeUnited States
 
dc.relation.ispartofPLoS One
 
dc.relation.referencesReferences in Scopus
 
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License
 
dc.subject.meshFungal Proteins - analysis - genetics - immunology - metabolism
 
dc.subject.meshImmunoassay - methods
 
dc.subject.meshMembrane Glycoproteins - analysis - genetics - immunology - metabolism
 
dc.subject.meshMycoses - blood - diagnosis - microbiology
 
dc.subject.meshPenicillium - genetics - growth and development - pathogenicity
 
dc.titleImmunoassays based on Penicillium marneffei Mp1p derived from Pichia pastoris expression system for diagnosis of penicilliosis
 
dc.typeArticle
 
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Author Affiliations
  1. Chinese National Human Genome Center at Shanghai
  2. The University of Hong Kong
  3. Zhujiang Hospital