Rapid Enzyme-Linked Immunosorbent Assay and Colloidal Gold Immunoassay for Kanamycin and Tobramycin in Swine Tissues

Chen, Yiqiang; Wang, Zhiqin; Wang, Zhanhui; Tang, Shusheng; Zhu, Yan; Xiao, X.

doi:10.1021/jf703602b

Cited by 115 publications

(73 citation statements)

References 42 publications

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“…These are often complex and require tedious sample pretreatment steps. The enzyme-linked immunosorbent assay (ELISA) has been proven for simple sample pretreatment with high sensitivity and selectivity (Chen et al 2008).…”

Section: Introductionmentioning

confidence: 99%

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

2015

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The present study was aimed to evaluate the feasibility to use chicken egg yolk antibodies (IgY) for the detection of Kanamycin (Kana) residues in food stuffs. Bovine serum albumin (BSA) was conjugated with Kana by carbodiimide method, and the laying chickens were immunized with Kana-BSA conjugate. PEG-6000 method was employed to extract IgY. The peak titer of anti-Kana IgY was 1:256,000 after the fifth booster immunization. The optimal dilution for anti-Kana IgY was 1:25,000 to obtain optical density (OD) of 1.0 at 2 μg/mL of OVA-Kana coating concentration. The indirect competitive ELISA (ic-ELISA) showed that the IC 50 value of anti-Kana IgY was 4.48 ng/ mL and the regression curve equation was y =−15.62x+ 60.17 (R 2 =0.98, n=3). The ic-ELISA showed a lower crossreactivity (less than 0.01 %) with other drugs (except gentamicin-1.91 %). Recoveries from milk, pork, and chicken powder samples were in the range of 82.02 to 98.20 %, with relative standard deviation lower than 5.13 %. Our results indicate that generated anti-Kana IgY can be used in routine screening analysis of Kana residues in food samples.

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Section: Introductionmentioning

confidence: 99%

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

2015

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“…In addition, no matrix effects due to hemolytic, lipemic, or icteric plasma matrices were observed. Multiple methods for the quantification of kanamycin have been published (8,9,11,(22)(23)(24)(25). Only one paper describes the use of an immunoassay for the determination of kanamycin where a tobramycin antibody was used as well (23).…”

Section: Discussionmentioning

confidence: 99%

Immunoassay Analysis of Kanamycin in Serum Using the Tobramycin Kit

Dijkstra

Voerman

Greijdanus

et al. 2016

Antimicrob Agents Chemother

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bKanamycin is one of the aminoglycosides used in the treatment of multidrug-resistant tuberculosis. Blood concentrations of kanamycin are predictive for the treatment efficacy and the occurrence of side effects, and dose adjustments can be needed to optimize therapy. However, an immunoassay method for the quantification of kanamycin is not commercially available. We modified the existing tobramycin immunoassay to analyze kanamycin. This modified method was tested in a concentration range of 0.3 to 80.0 mg/liter for inaccuracy and imprecision. In addition, the analytical results of the immunoassay method were compared to those obtained by a liquid chromatography-tandem mass spectrometry (LC-MS/MS) analytical method using Passing and Bablok regression. Within-day imprecision varied from 2.3 to 13.3%, and between-day imprecision ranged from 0.0 to 11.3%. The inaccuracy ranged from ؊5.2 to 7.6%. No significant cross-reactivity with other antimicrobials and antiviral agents was observed. The results of the modified immunoassay method were comparable with the LC-MS/MS analytical outcome. This new immunoassay method enables laboratories to perform therapeutic drug monitoring of kanamycin without the need for complex and expensive LC-MS/MS equipment.T uberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis. Unfortunately, resistance to the two firstline drugs in TB treatment, isoniazid and rifampin, is emerging (1). Treatment regimens for multidrug-resistant TB (MDR-TB) include a quinolone and an injectable: amikacin, kanamycin, or capreomycin (2). Unfortunately, the use of aminoglycosides comes with toxic effects, such as renal failure and irreversible hearing loss (3).Recently, a study with MDR-TB patients showed that the toxicity of aminoglycosides was correlated with the cumulative area under the curve (4). For efficacy, it is well recognized that the top serum concentration (C max ) is related to the efficacy of aminoglycosides (5). In addition, pharmacokinetic (PK) guided dosing reduced the daily dose of aminoglycosides, with excellent treatment outcome (6). This indicates that serum concentration monitoring may be of added value in the treatment of tuberculosis. The targeted peak concentration for kanamycin in MDR-TB is 20 to 30 mg/liter (7), while the trough concentration should be as low as reasonably achievable in order to prevent toxicity.Although analytical methods to quantify the serum or plasma concentrations of amikacin and kanamycin using liquid chromatography-tandem mass spectrometry (LC-MS/MS) (8-11) have been described, this technique is commonly not available in most developing countries (12). However, in contrast to the case for amikacin, no analytical immunoassay kit to analyze kanamycin is commercially available.Kanamycin is more structurally related to tobramycin than to amikacin (Fig. 1). The product insert of the tobramycin immunoassay kit, based on the enzyme multiplied immunoassay technique (EMIT), indicates cross-reactivity for kanamycin (13). We therefore explored the...

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“…Portions of the cell fusion procedures and cloning conditions were described previously by Köhler and Milstein [18] with further modifications by Chen et al [19]. Briefly, NS0 myeloma cells were passed through a medium containing 8-azaguanine and then grown for 4-5 d at 37°C in a 5% CO 2 atmosphere.…”

Section: Fusion Of Myeloma and Spleen Cellsmentioning

confidence: 99%

Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue

et al. 2011

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has been illegally used in horse racing to boost physical performance, and in animal husbandry to accelerate weight gain. To monitor the abuse of NT, our goal was to develop a commercial enzyme linked immunosorbent assay (ELISA) kit. For this purpose, hybridomas were prepared by fusing NS0 mouse myeloma cells with splenocytes isolated from immunized BALB/c mouse. Noncompetitive and competitive indirect ELISA were used to screen positive cell clones. To optimize the indirect competitive ELISA (icELISA) method, various methanol concentrations in assay buffer were evaluated. Matrix effects in urine and spiking test were also investigated. Finally, five hybridoma cell lines named NT-1, NT-2, NT-3, NT-4 and NT-5 were screened out. The corresponding monoclonal antibodies (mAbs) were of the IgG 1 isotype with a k light chain, and the antibody affinity of all mAbs were between 2.6×10 9 and 4.7×10 9 L/mol. The titer and IC 50 values of purified ascites were in the range of 0.64×10 5 −2.56×10 5 and 0.55-1.0 ng/mL, respectively. Based on the NT-1 hybridoma, a heterologous icELISA method was developed for the quantitative detection of NT in cattle urine. The dynamic range was from 0.004 to 85.8 ng/mL, with a detection limit for the assay and IC 50 values of 0.002 and 0.55 ng/mL, respectively. Except for a high cross-reactivity (62%) to α-NT, negligible cross-reactivity to other compounds was observed. After optimization, 10% of methanol was used in the assay buffer, and a 20-fold dilution in cattle urine gave an inhibition curve almost the same as that in phosphate buffered saline. The correlation coefficient between the established icELISA and LC-MS/MS method was 0.9871. The results showed that the established heterologous icELISA method provides an excellent alternative for the detection of NT residues in food producing animals. 19-nortestosterone

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Rapid Enzyme-Linked Immunosorbent Assay and Colloidal Gold Immunoassay for Kanamycin and Tobramycin in Swine Tissues

Cited by 115 publications

References 42 publications

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

Preparation of Artificial Antigen and Development of IgY-Based Indirect Competitive ELISA for the Detection of Kanamycin Residues

Immunoassay Analysis of Kanamycin in Serum Using the Tobramycin Kit

Preparation of monoclonal antibody based indirect competitive ELISA for detecting 19-nortestosterone residue

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