Diagnosis of tularemia using piezoelectric biosensor technology |
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Authors: | Miroslav Pohanka,Oto Pavli&scaron |
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Affiliation: | a Center of Biological Defense, 56166 Těchonín, Czech Republic b Department of Biochemistry, Masaryk University, Kotlá?ská 2, 61137 Brno, Czech Republic |
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Abstract: | A piezoelectric immunosensor for indirect diagnosis of tularemic infection in mouse serum was developed. Francisella tularensis LVS antigen was covalently immobilized on the sensing surface using cystamine and glutaraldehyde for activation and modification of the gold electrode. The normal mouse serum (NMS) and serum prepared from mice immunized by Escherichia coli were used as negative controls providing signal of 28 Hz during a 5 min interaction. The tularemic infectious (immunized) mouse serum (IMS) as sample resulted in the signal above 75 Hz (fifth day after infection). The control sensor containing bovine serum albumin as sensing element provided a signal below 5 Hz with NMS as well IMS. The effects of dilution degree and purification of sera were tested. To improve resolution of the method, sample pretreatment steps such as precipitation with ammonium sulphate and immunoglobulin extraction on CBind™ L and MEP HyperCel columns were tested. R.S.D. of measurements was 2.3% for NMS and 2.4% for IMS, respectively. The developed method allows to indicate the presence of anti-tularemic antibodies shortly (1-3 days) after infection, one analysis is completed in 10 min. |
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Keywords: | Piezoelectric immunosensor Francisella tularensis Serum Biowarfare agent |
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