Rapid and sensitive enzyme-linked immunosorbent assay and immunochromatographic assay for the detection of chlortetracycline residues in edible animal tissues

Abstract

Two rapid and sensitive enzyme-linked immunosorbent assays (ELISA) and an immunochromatographic assay (ICA) for the detection of chlortetracycline (CTC) residues in edible animal tissues were developed based on a monoclonal antibody (MAb) produced by using the chlortetracycline-bovine serum albumin (CTC-BSA) conjugate as the immunogen. A total of 50% inhibiting concentration (IC(50)) of the modified ELISA was 0.66 ng ml(-1) and the recoveries from spiked chicken muscle and liver were 78.8-92.2% and 80.3-90.2%, respectively. The corresponding coefficient variations (CVs) were 3.2-9.5% and 6.5-10.2%. The detection limit was 0.06 ng g(-1) in chicken muscle and 0.07 ng g(-1) in liver. However, the detection limit of ICA was 0.12 ng ml(-1), and the recoveries in negative samples spiked at concentrations of 10, 50 and 100 ng g(-1) ranged from 79.0% to 88.6% for muscle samples and from 75.2% to 87.0% for liver samples. The cut-off values for the test lines were 80 ng g(-1) and the analysis can be completed within 5-10 min. Comparisons with an HPLC method were performed by testing 200 swine muscle samples and chicken muscle samples from local markets, and an agreement rate of 99.5% was obtained between the three methods.

Production of a monoclonal antibody against oxytetracycline and its application for oxytetracycline residue detection in shrimp

“…Because the properties of PAbs vary among batches according to the immune responses of different immunized animals, PAbs are less suitable for long-term application than MAbs (Hock et al, 1995). In addition, MAbs against CTC and DC have been reported (Le et al, 2011a;2011b). However, these MAbs had low cross-reactivity to OTC.…”

“…Importantly, an immunoassay does not require a complicated step to clean up samples (Cháfer-Pericása et al, 2010). Previously, established ELISA methods have been based on a polyclonal antibody (PAb) for doxycycline (DC) detection (Le et al, 2009) and a monoclonal antibody (MAb) for chlortetracycline (CTC) detection (Le et al, 2011b). The use of a MAb is preferable due to its uniformity and unlimited production with unchanged properties.…”

Methods for the separation, preconcentration, and determination of tetracycline antibiotics

“…HPLC makes it possible to determine tetracycline at a level of hundredths of μg/mL. Food products and feed IC [46] Meat HPLC [15], [19], [21][22][23], [27], [51] SP [85] IC [16][17][18], [20], [27] VA [24], [31] SP [85] Environmental samples VA [24] Water HPLC [72], [86], [89], [93], [94], [100], [ Eggs HPLC [27], [32], [53], [68], [134] FL [117], [119], [156] IC [18], [27] …”

“…Liquid extraction from solid matrices seems the most common method for the separation of tetracy clines from the solid samples of food products. This method is used for the extraction of tetracycline anti biotics from meat [15][16][17][18][19][20][21][22][23][24][25][26][27], fish [27][28][29][30][31], eggs [18,27,32], and honey [33][34][35][36][37][38][39], and it is performed as follows: A weighed portion of a thoroughly ground solid sam ple is placed in a vessel for agitation, and a selected sol vent is added; the contents are stirred for a specified time (from several minutes to several hours). The phases are separated by filtration.…”

“…Enzyme linked immunosorbent assay (ELISA) is the most fre quently used method. With the use of ELISA, the tet racycline antibiotics were determined in meat prod ucts [16][17][18]20], fish [30,46], milk [20,135,136,138,[141][142][143][144][145], eggs [18,27], and honey [38,135,138]. Electrochemical immunoenzyme assay was used for the determination of tetracycline antibiotics on the biotin-avidin conjugated CdS and PbS nanoclusters [135] and graphene nanosheets covered with Pt nano particles [136].…”