Detection of Leptospira in urine using anti-Leptospira-coated gold nanoparticles

Abstract

Serological assays for antibody detection have been widely used for Leptospirosis diagnosis. However, antibody is usually undetectable during the first week after infection. Detection of Leptospira DNA can be done by PCR but this technique requires special equipments and the cost is still relatively high. Here we demonstrate that gold nanoparticles can be used to facilitate Leptospira detection. Gold nanoparticles were coated with rabbit antibody specific to Leptospira interrogans serovar Bratislava and these coated particles were used to detect Leptospira in urine. Agglutination of gold particles indicated the presence of Leptospira in samples tested. The sensitivity of detection was 10 leptospires/ml. No agglutination was observed when anti-Leptospira-coated particles were tested with urine containing the organisms commonly found in urine such as Klebsiella pneumoniae, Escherichia coli and Enterococcus faecalis. This assay is very easy to perform and results could be observed with the naked eyes. Fresh or frozen urine samples could be used. The stability of antibody-coated particles was at least 2 months when kept at 4°C. In conclusion, we have demonstrated that the technique using antibody-coated gold nanoparticles is a promising tool for further validation as a rapid assay for Leptospirosis diagnosis.