TY - JOUR
T1 - Conservation of mannan synthesis in fungi of the Zygomycota and Ascomycota reveals a broad diagnostic target
AU - Burnham-Marusich, Amanda R.
AU - Hubbard, Breeana
AU - Kvam, Alexander J.
AU - Gates-Hollingsworth, Marcellene
AU - Green, Heather R.
AU - Soukup, Eric
AU - Limper, Andrew H.
AU - Kozel, Thomas R.
N1 - Funding Information:
This work was supported by Public Health Service grants R33 AI085548 (to T.R.K.), R41 AI102311 (subcontract to T.R.K.), R41 AI108114 (to T.R.K.), R43 EB023408 (to A.R.B.-M.), and R01 HL62150 (to A.H.L.) from the National Institutes of Health. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Funding Information:
We thank Shouhua Wang, Nevada Department of Agriculture, Jamie Voyles, Department of Biology, University of Nevada-Reno, Gary Blomquist, Department of Biochemistry and Molecular Biology, University of Nevada-Reno, and William Courchesne, Department of Microbiology & Immunology, University of Nevada-Reno, for providing cultures and/or fungus-infected tissue. We thank Dillon Petretti and Raghavi Anand for assistance with experiments. This work was supported by Public Health Service grants R33 AI085548 (to T.R.K.), R41 AI102311 (subcontract to T.R.K.), R41 AI108114 (to T.R.K.), R43 EB023408 (to A.R.B.-M.), and R01 HL62150 (to A.H.L.) from the National Institutes of Health. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. T.R.K. has an equity interest in DxDiscovery, a university startup firm focused on early-stage diagnostics development. A.R.B.-M. is also a principal scientist at DxDiscovery
Publisher Copyright:
© 2018 Burnham-Marusich et al.
PY - 2018/5/1
Y1 - 2018/5/1
N2 - Ascomycetes and zygomycetes account for the majority of (i) fungi responsible for cutaneous, subcutaneous, and invasive human fungal infections, (ii) plant fungal pathogens, (iii) fungi that threaten global biodiversity, (iv) fungal agents of agricultural spoilage, and (v) fungi in water-damaged buildings. Rapid recognition of fungal infection (or contamination) enables early treatment (or remediation). A bioinformatics search found homologues of Saccharomyces cerevisiae Mnn9p present in members of the Zygomycota and Ascomycota phyla and absent in members of the Chytridiomycota and Basidiomycota. Mnn9p is a component of the yeast mannan polymerization complex and is necessary for α-1,6 mannan production. A monoclonal antibody (2DA6) was produced that was reactive with purified mannans of Mucor, Rhizopus, Aspergillus, Fusarium, and Candida species. Experimentation using a 2DA6 antigen capture enzyme-linked immunosorbent assay (ELISA) and extracts of fungi from the four phyla found agreement between the presence or absence of Mnn9p homologues and production or lack of production of mannan reactive with 2DA6. Studies of cell extracts from yeast mannan mutants identified α-1,6 mannan as the epitope recognized by 2DA6. To translate this finding into a point-of-use diagnostic, a 2DA6 lateral flow immunoassay was constructed that detected mannan in (i) extracts of dermatophytes and fungi that produce traumarelated infection and (ii) tissue from plants infected with Grosmannia clavigera or Sclerotium cepivorum. These studies (i) revealed that the conservation of α-1,6-linked mannan in fungi of the Zygomycota and Ascomycota can be exploited as a broad diagnostic target and (ii) have provided a means to detect that target in an immunoassay platform that is well suited for clinic or field use.
AB - Ascomycetes and zygomycetes account for the majority of (i) fungi responsible for cutaneous, subcutaneous, and invasive human fungal infections, (ii) plant fungal pathogens, (iii) fungi that threaten global biodiversity, (iv) fungal agents of agricultural spoilage, and (v) fungi in water-damaged buildings. Rapid recognition of fungal infection (or contamination) enables early treatment (or remediation). A bioinformatics search found homologues of Saccharomyces cerevisiae Mnn9p present in members of the Zygomycota and Ascomycota phyla and absent in members of the Chytridiomycota and Basidiomycota. Mnn9p is a component of the yeast mannan polymerization complex and is necessary for α-1,6 mannan production. A monoclonal antibody (2DA6) was produced that was reactive with purified mannans of Mucor, Rhizopus, Aspergillus, Fusarium, and Candida species. Experimentation using a 2DA6 antigen capture enzyme-linked immunosorbent assay (ELISA) and extracts of fungi from the four phyla found agreement between the presence or absence of Mnn9p homologues and production or lack of production of mannan reactive with 2DA6. Studies of cell extracts from yeast mannan mutants identified α-1,6 mannan as the epitope recognized by 2DA6. To translate this finding into a point-of-use diagnostic, a 2DA6 lateral flow immunoassay was constructed that detected mannan in (i) extracts of dermatophytes and fungi that produce traumarelated infection and (ii) tissue from plants infected with Grosmannia clavigera or Sclerotium cepivorum. These studies (i) revealed that the conservation of α-1,6-linked mannan in fungi of the Zygomycota and Ascomycota can be exploited as a broad diagnostic target and (ii) have provided a means to detect that target in an immunoassay platform that is well suited for clinic or field use.
KW - Diagnostics
KW - Immunodetection
KW - Invasive fungal infection
KW - Lateral flow immunoassay
KW - Mannan
KW - Mnn9
KW - Point of care
UR - http://www.scopus.com/inward/record.url?scp=85048227733&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85048227733&partnerID=8YFLogxK
U2 - 10.1128/mSphere.00094-18
DO - 10.1128/mSphere.00094-18
M3 - Article
C2 - 29720523
AN - SCOPUS:85048227733
SN - 2379-5042
VL - 3
JO - mSphere
JF - mSphere
IS - 3
M1 - e00094-18
ER -