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Sensitive, fast, and specific immunoassays for methyltestosterone detection.

Kong N, Song S, Peng J, Liu L, Kuang H, Xu C - Sensors (Basel) (2015)

Bottom Line: Based on icELISA results, the recovery rate of MT in animal feed was 82.4%-100.6%.The results were in accordance with those obtained by gas chromatography-mass spectrometry.Therefore, these immunoassays are useful and fast tools for MT residue detection in animal feed.

View Article: PubMed Central - PubMed

Affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China. kongxiyangsucc@126.com.

ABSTRACT
An indirect competitive enzyme-linked immunosorbent assay (icELISA) and an immunochromatographic strip assay using a highly specific monoclonal antibody, were developed to detect methyltestosterone (MT) residues in animal feed. The optimized icELISA had a half-inhibition concentration value of 0.26 ng/mL and a limit of detection value of 0.045 ng/mL. There was no cross-reactivity with eight analogues, revealing high specificity for MT. Based on icELISA results, the recovery rate of MT in animal feed was 82.4%-100.6%. The results were in accordance with those obtained by gas chromatography-mass spectrometry. The developed immunochromatographic strip assay, as the first report for MT detection, had a visual cut-off value of 1 ng/mL in PBS, 2.5 ng/g in fish feed, and 2.5 ng/g in pig feed. Therefore, these immunoassays are useful and fast tools for MT residue detection in animal feed.

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Optimized inhibition standard curve for MT analysis by icELISA. Error bar was calculated according to three repeats at each concentration.
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sensors-15-10059-f002: Optimized inhibition standard curve for MT analysis by icELISA. Error bar was calculated according to three repeats at each concentration.

Mentions: In conclusion, the MT standard was diluted with PBS (10 mM, pH 7.4) and analyzed by icELISA. The standard curve was established by plotting the B/B0 (B was the absorbance at each concentration of MT and B0 was the absorbance in the absence of MT) against the concentration of MT (Figure 2). The standard curve equation was y = 0.081 + (0.999 − 0.081)/(1 + (x/0.229)1.39), the linear regression correlation coefficient (R2) was 0.999, the IC50 value was 0.26 ng/mL, and the limit of detection (LOD, concentration calculated as IC10) was 0.045 ng/mL.


Sensitive, fast, and specific immunoassays for methyltestosterone detection.

Kong N, Song S, Peng J, Liu L, Kuang H, Xu C - Sensors (Basel) (2015)

Optimized inhibition standard curve for MT analysis by icELISA. Error bar was calculated according to three repeats at each concentration.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4481955&req=5

sensors-15-10059-f002: Optimized inhibition standard curve for MT analysis by icELISA. Error bar was calculated according to three repeats at each concentration.
Mentions: In conclusion, the MT standard was diluted with PBS (10 mM, pH 7.4) and analyzed by icELISA. The standard curve was established by plotting the B/B0 (B was the absorbance at each concentration of MT and B0 was the absorbance in the absence of MT) against the concentration of MT (Figure 2). The standard curve equation was y = 0.081 + (0.999 − 0.081)/(1 + (x/0.229)1.39), the linear regression correlation coefficient (R2) was 0.999, the IC50 value was 0.26 ng/mL, and the limit of detection (LOD, concentration calculated as IC10) was 0.045 ng/mL.

Bottom Line: Based on icELISA results, the recovery rate of MT in animal feed was 82.4%-100.6%.The results were in accordance with those obtained by gas chromatography-mass spectrometry.Therefore, these immunoassays are useful and fast tools for MT residue detection in animal feed.

View Article: PubMed Central - PubMed

Affiliation: State Key Lab of Food Science and Technology, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China. kongxiyangsucc@126.com.

ABSTRACT
An indirect competitive enzyme-linked immunosorbent assay (icELISA) and an immunochromatographic strip assay using a highly specific monoclonal antibody, were developed to detect methyltestosterone (MT) residues in animal feed. The optimized icELISA had a half-inhibition concentration value of 0.26 ng/mL and a limit of detection value of 0.045 ng/mL. There was no cross-reactivity with eight analogues, revealing high specificity for MT. Based on icELISA results, the recovery rate of MT in animal feed was 82.4%-100.6%. The results were in accordance with those obtained by gas chromatography-mass spectrometry. The developed immunochromatographic strip assay, as the first report for MT detection, had a visual cut-off value of 1 ng/mL in PBS, 2.5 ng/g in fish feed, and 2.5 ng/g in pig feed. Therefore, these immunoassays are useful and fast tools for MT residue detection in animal feed.

Show MeSH