Preparation of immunomagnetic beads coupled with a rhodamine hydrazine immunosensor for the detection of Mycobacterium avium subspecies paratuberculosis in bovine feces, milk, and colostrum
Autor: | Peter D. Constable, M. Nouri, M. Khosravi, N. Mosavari, A. Mohammadi |
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Rok vydání: | 2021 |
Předmět: |
animal diseases
Cattle Diseases Hydrazone Paratuberculosis Biosensing Techniques Rhodamine Feces 03 medical and health sciences chemistry.chemical_compound fluids and secretions Pregnancy Genetics medicine Animals 030304 developmental biology Carbodiimide Immunoassay chemistry.chemical_classification Detection limit 0303 health sciences Chromatography biology Rhodamines Chromogenic Colostrum 0402 animal and dairy science 04 agricultural and veterinary sciences biology.organism_classification medicine.disease 040201 dairy & animal science Mycobacterium avium subspecies paratuberculosis Mycobacterium avium subsp. paratuberculosis Hydrazines Milk chemistry Cattle Female Animal Science and Zoology Food Science |
Zdroj: | Journal of Dairy Science. 104:6944-6960 |
ISSN: | 0022-0302 |
Popis: | The aim of this study was to develop and evaluate a method for detecting Mycobacterium avium ssp. paratuberculosis (MAP) bacteria in bovine fecal, milk, and colostrum samples using immunomagnetic beads (IMB) and a rhodamine hydrazone immunosensor. Immunomagnetic beads were prepared by using purified antibodies from hyperimmunized sera that were coupled to Fe nanoparticles with diethylene triamine pentaacetic acid (DTPA) or ethyl (dimethyl aminopropyl) carbodiimide (EDC)-N-hydroxy succinimide (NHS) as linkers. Rhodamine hydrazone particles were synthesized and coupled to IgY anti-MAP antibodies using DTPA or EDC-NHS linkers. Separation efficiency of the IMB was tested on bovine fecal, milk, and colostrum samples experimentally contaminated with MAP. The studied methods were evaluated on their ability to detect MAP and separate bacteria in complex mediums. The ELISA results indicated 95% efficacy in antibody coupling to IMB, with the DTPA-IMB method being more efficient than the EDC-NHS-IMB method. By using the DTPA-IMB method, MAP bacteria were successfully recovered from fecal, milk, and colostrum samples. The DTPA-IMB method used in combination with the rhodamine hydrazone immunosensor had a limit of detection equal to 30 and 30,000 MAP cells/mL using chromogenic and fluorescent properties, respectively. Combining the DTPA-IMB separation method with the rhodamine hydrazone immunosensor provides a fast, sensitive, and cost-beneficial method for detecting MAP in bovine feces, milk, and colostrum. |
Databáze: | OpenAIRE |
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