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Steroids. 2017 Oct;126:57-65. doi: 10.1016/j.steroids.2017.07.005. Epub 2017 Jul 14.

A robust high-throughput fungal biosensor assay for the detection of estrogen activity.

Author information

1
Research Platform Bioactive Microbial Metabolites (BiMM), Bioresources and Technologies Campus Tulln, Konrad Lorenz Straße 24, 3430 Tulln, Austria; Institute of Milk Hygiene, Milk Technology and Food Science, Department of Farm Animal and Veterinary Public Health, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
2
University Clinic for Ruminants, Clinical Unit for Herd Health Management in Ruminants, Department for Farm Animals and Veterinary Public Health, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria. Electronic address: Karen.Wagener@vetmeduni.ac.at.
3
Research Platform Bioactive Microbial Metabolites (BiMM), Bioresources and Technologies Campus Tulln, Konrad Lorenz Straße 24, 3430 Tulln, Austria.
4
University Clinic for Ruminants, Clinical Unit for Herd Health Management in Ruminants, Department for Farm Animals and Veterinary Public Health, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
5
Institute of Physiology, Pathophysiology and Biophysics, Department of Biomedical Sciences, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
6
Institute of Milk Hygiene, Milk Technology and Food Science, Department of Farm Animal and Veterinary Public Health, Vetmeduni Vienna, Veterinärplatz 1, 1210 Vienna, Austria.
7
Research Platform Bioactive Microbial Metabolites (BiMM), Bioresources and Technologies Campus Tulln, Konrad Lorenz Straße 24, 3430 Tulln, Austria; Fungal Genetics and Genomics Unit, Department of Applied Genetics and Cell Biology, BOKU University of Natural Resources and Life Sciences Vienna, Konrad Lorenz Straße 24, 3430 Tulln, Austria.

Abstract

Estrogenic active compounds are present in a variety of sources and may alter biological functions in vertebrates. Therefore, it is crucial to develop innovative analytical systems that allow us to screen a broad spectrum of matrices and deliver fast and reliable results. We present the adaptation and validation of a fungal biosensor for the detection of estrogen activity in cow derived samples and tested the clinical applicability for pregnancy diagnosis in 140 mares and 120 cows. As biosensor we used a previously engineered genetically modified strain of the filamentous fungus Aspergillus nidulans, which contains the human estrogen receptor alpha and a reporter construct, in which β-galactosidase gene expression is controlled by an estrogen-responsive-element. The estrogen response of the fungal biosensor was validated with blood, urine, feces, milk and saliva. All matrices were screened for estrogenic activity prior to and after chemical extraction and the results were compared to an enzyme immunoassay (EIA). The biosensor showed consistent results in milk, urine and feces, which were comparable to those of the EIA. In contrast to the EIA, no sample pre-treatment by chemical extraction was needed. For 17β-estradiol, the biosensor showed a limit of detection of 1ng/L. The validation of the biosensor for pregnancy diagnosis revealed a specificity of 100% and a sensitivity of more than 97%. In conclusion, we developed and validated a highly robust fungal biosensor for detection of estrogen activity, which is highly sensitive and economic as it allows analyzing in high-throughput formats without the necessity for organic solvents.

KEYWORDS:

Aspergillus nidulans; Bioassay; Estrogen activity; Green chemistry; Pregnancy diagnosis

PMID:
28712952
DOI:
10.1016/j.steroids.2017.07.005
[Indexed for MEDLINE]

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