Development of immunocolloidal strip for rapid detection of pyrimethanil

Abstract

In this study, we prepared a monoclonal antibody against pyrimethanil, for which we first derived its hapten according to the structural formula of pyrimethanil; we used indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) to screen positive mice for cell fusion; finally, we developed five cell lines (1A5, 1E7, 3A6, 3H12 and 5D4) that produced monoclonal antibodies to pyrimethanil. The monoclonal antibody produced by each cell was detected by ic-ELISA, and the monoclonal antibody produced by the 1E7 cell strain was found to be the most sensitive to pyrimethanil. The IC50 value of this antibody was 4 ng/ml. After using this monoclonal antibody, i.e. 1E7, to perform immunocolloidal gold strip treatment on cucumber samples, the PBS and the cucumber samples cutoff value of Immunochromatographic strips (ICS) was found to be 50 ng/ml; We conclude that ICS can rapidly detect pyrimethanil in cucumber samples.

Development and validation of an immunochromatography test strip for rapid detection of pyrimethanil residues

“…The whole procedure was simplified, and the assay time was shorten comparing with the ic-ELISA we developed before (Cao et al, 2019). For qualitative evaluation, the cutoff values of ICTS for pyrimethanil was 40 ng/mL The cut-off value of the proposed strip was lower than that of strip (cut-off value of 50 ng/mL) in the previous literature (Chen et al, 2019). By scanning the ROD of the test line, this ICTS could be quantitatively analysed by the strip reader.…”

“…These analytical methods are sensitive and accurate, whilst costly equipment, skilled personnel high cost, and unsuitable for on-site detection are their drawbacks. Consequently, rapid immunoassay methods such as ELISA indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) (Cao, Shi, Le, Tang, & Xie, 2019;Chen et al, 2019;Mercader, Esteve-Turrillas, Agullo, Abad-Somovilla, & Abad-Fuentes, 2012) and colloidal gold immunochromatography test strip (ICTS) (Chen et al, 2019) have been widely used for the rapid detection of pyrimethanil residue. These methods and their reported detection sensitivities are listed in Table 1.…”

Rapid detection of rifampicin in fish using immunochromatographic strips

“…The caprylic acid-ammonium sulphate precipitation method was used to purify the ascites. The purified mAbs were stored at −20°C (Chen et al, 2019).…”

Development of an immunocolloidal strip for rapid detection of picoxystrobin

“…After the solution was layered, the lower layer containing the red gold-labeled mAb was collected and washed with a 0.02 M phosphate buffer solution (5% sucrose, 1% BSA and 0.5% PEG 6000). The combined product was reconstituted to 1 mL with gold-labeled resuspension buffer (0.02 M PBS, 5% sucrose, 2% sorbitol, 1% mannitol, 0.1% PEG, 0.1% Tween and 0.04% NaN 3 ) and stored at 4°C (Chen et al, 2019).…”