Limits...
Development of a Monoclonal Antibody-Based icELISA for the Detection of Ustiloxin B in Rice False Smut Balls and Rice Grains.

Fu X, Wang A, Wang X, Lin F, He L, Lai D, Liu Y, Li QX, Zhou L, Wang B - Toxins (Basel) (2015)

Bottom Line: The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B.Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g.Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC) demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

View Article: PubMed Central - PubMed

Affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China. xiaoxiaofu@cau.edu.cn.

ABSTRACT
Rice false smut is an emerging and economically-important rice disease caused by infection by the fungal pathogen Villosiclava virens. Ustiloxin B is an antimitotic cyclopeptide mycotoxin isolated from the rice false smut balls that formed in the pathogen-infected rice spikelets. A monoclonal antibody (mAb) designated as mAb 1B5A10 was generated with ustiloxin B-ovalbumin conjugate. A highly-sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed. The median inhibitory concentration (IC50) of the icELISA was 18.0 ng/mL for the detection of ustiloxin B; the limit of detection was 0.6 ng/mL, and the calibration range was from 2.5 to 107.4 ng/mL. The LOD/LOQ values of the developed ELISA used for the determination of ustiloxin B in rice false smut balls and rice grains were 12/50 μg/g and 30/125 ng/g, respectively. The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B. Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g. Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC) demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

No MeSH data available.


Related in: MedlinePlus

Inhibition curve of ustiloxin B in indirect competitive ELISA (icELISA) format based on mAb IB5A10 (each value represents the mean of triplicate ± standard deviations; B0 and B are the absorbance values at 492 nm in the absence and presence of ustiloxin B, respectively).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4591656&req=5

toxins-07-03481-f002: Inhibition curve of ustiloxin B in indirect competitive ELISA (icELISA) format based on mAb IB5A10 (each value represents the mean of triplicate ± standard deviations; B0 and B are the absorbance values at 492 nm in the absence and presence of ustiloxin B, respectively).

Mentions: The icELISA measurements were conducted with a series of concentrations (0, 1.17, 2.34, 4.69, 9.38, 18.75, 37.5, 75, 150, 300 ng/mL) of ustiloxin B dissolved in PBSTG under the optimal conditions. A representative inhibition curve (Figure 2) for ustiloxin B generated by icELISA based on mAb IB5A10 was established. The median inhibitory concentration (IC50) of the icELISA was 18.0 ng/mL. The limit of detection was 0.6 ng/mL (10% inhibition). The calibration range, based on 20% to 80% of inhibition of the binding of mAb 1B5A10 to the immobilized hapten-BSA, was from 2.5 to 107.4 ng/mL.


Development of a Monoclonal Antibody-Based icELISA for the Detection of Ustiloxin B in Rice False Smut Balls and Rice Grains.

Fu X, Wang A, Wang X, Lin F, He L, Lai D, Liu Y, Li QX, Zhou L, Wang B - Toxins (Basel) (2015)

Inhibition curve of ustiloxin B in indirect competitive ELISA (icELISA) format based on mAb IB5A10 (each value represents the mean of triplicate ± standard deviations; B0 and B are the absorbance values at 492 nm in the absence and presence of ustiloxin B, respectively).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591656&req=5

toxins-07-03481-f002: Inhibition curve of ustiloxin B in indirect competitive ELISA (icELISA) format based on mAb IB5A10 (each value represents the mean of triplicate ± standard deviations; B0 and B are the absorbance values at 492 nm in the absence and presence of ustiloxin B, respectively).
Mentions: The icELISA measurements were conducted with a series of concentrations (0, 1.17, 2.34, 4.69, 9.38, 18.75, 37.5, 75, 150, 300 ng/mL) of ustiloxin B dissolved in PBSTG under the optimal conditions. A representative inhibition curve (Figure 2) for ustiloxin B generated by icELISA based on mAb IB5A10 was established. The median inhibitory concentration (IC50) of the icELISA was 18.0 ng/mL. The limit of detection was 0.6 ng/mL (10% inhibition). The calibration range, based on 20% to 80% of inhibition of the binding of mAb 1B5A10 to the immobilized hapten-BSA, was from 2.5 to 107.4 ng/mL.

Bottom Line: The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B.Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g.Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC) demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

View Article: PubMed Central - PubMed

Affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China. xiaoxiaofu@cau.edu.cn.

ABSTRACT
Rice false smut is an emerging and economically-important rice disease caused by infection by the fungal pathogen Villosiclava virens. Ustiloxin B is an antimitotic cyclopeptide mycotoxin isolated from the rice false smut balls that formed in the pathogen-infected rice spikelets. A monoclonal antibody (mAb) designated as mAb 1B5A10 was generated with ustiloxin B-ovalbumin conjugate. A highly-sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) was then developed. The median inhibitory concentration (IC50) of the icELISA was 18.0 ng/mL for the detection of ustiloxin B; the limit of detection was 0.6 ng/mL, and the calibration range was from 2.5 to 107.4 ng/mL. The LOD/LOQ values of the developed ELISA used for the determination of ustiloxin B in rice false smut balls and rice grains were 12/50 μg/g and 30/125 ng/g, respectively. The mAb 1B5A10 cross-reacted with ustiloxin A at 13.9% relative to ustiloxin B. Average recoveries of ustiloxin B ranged from 91.3% to 105.1% for rice false smut balls at spiking levels of 0.2 to 3.2 mg/g and from 92.6% to 103.5% for rice grains at spiking levels of 100 to 5000 ng/g. Comparison of ustiloxin B content in rice false smut balls and rice grains detected by both icELISA and high performance liquid chromatography (HPLC) demonstrated that the developed icELISA can be employed as an effective and accurate method for the detection of ustiloxin B in rice false smut balls, as well as rice food and feed samples.

No MeSH data available.


Related in: MedlinePlus