Development of an immunochromatographic lateral flow device for rapid diagnosis of Vibrio cholerae O1 serotype Ogawa

Vibrio cholerae and Cholera biotypes

“…Chen et al (2014) developed a new monoclonal antibody (McAb) pair, named IXiao3G6 and IXiao1D9, which is specifically against V. cholerae O1 serotype Ogawa. In addition, they developed immunochromatographic lateral flow device (LFD) using this McAb pair for the highly specific and rapid (within 5 min) detection of Ogawa.…”

The epitope analysis of an antibody specifically against Vibrio cholerae O1 Ogawa by phage library study

“…To test whether the identified mimic peptide can inhibit the interaction between McAb IXiao 3 G 6 and Ogawa, the 10 μg/mL McAb IXiao 3 G 6 coated plates were incubated with 50 μL inactivated bacteria (V. cholerae O1 Ogawa and Inaba, V. cholerae O139, E. coli, S. typhi, S. sonnei, V. fluvialis, and A. caviae) (refer to Section 2.1 for details), or alternatively their corresponding LPS extracts (prepared as previously described (Chen et al, 2014)) at different concentrations (10, 5, 2.5, 1.25, 0.625, 0.31 μg/mL) diluted in PBS for 10 min at 37°C. After being washed in TBST for 5 times, plates were incubated with either 50 μL P16 phage clone (1.0 × 10 7 pfu) or P17 phage clone (1.0 × 10 7 pfu) respectively, followed by anti-M13-HRP incubation and color development.…”

“…The Ph.D.™-7 Phage Display Peptide Library was purchased from New England Biolabs (New England Biolabs (Beijing) Ltd., Beijing, China). The 96-well Microtiter™ plates (polystyrene, flat, Thermo Scientific, China) were firstly coated with 100 μL McAb IXiao 3 G 6 (100 μg/mL) (Chen et al, 2014), diluted in 0.1 M NaHCO 3 , and incubated overnight at 4°C. Plates were then incubated for 2 h with 200 μL blocking buffer 0.1 M NaHCO 3 /1% BSA at room temperature and washed three times in Tris-Buffered Saline plus 0.05% Tween 20 (TBST).…”

“…The inactivated bacteria were quantified as protein in this study, and the protein content of all preparations was determined by standard BCA assay. After each bacterial strain was biochemically characterized and evaluated as previously described (Chen et al, 2014). The Ph.D.™-7 Phage Display Peptide Library was purchased from New England Biolabs (New England Biolabs (Beijing) Ltd., Beijing, China).…”

“…The present study intended to employ a monoclonal antibody we previously generated, IXiao 3 G 6 (Chen et al, 2014), which is directed against LPS of V. cholerae O1 Ogawa with competitive sensitivity and specificity, to identify the specific mimic peptides with high avidity through biopanning using phage random 7 peptide library.…”