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KMID : 1143420230160441491
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Public Health Weekly Report
2023 Volume.16 No. 44 p.1491 ~ p.1503
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Development of a Laboratory Test for Botulism Using Sandwich ELISA
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Choi Eun-Sun
Kim So-Hyeon
Jeon Jun-Ho
Yi Hwa-Jung
Chung Yoon-Seok
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Abstract
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Botulism is a flaccid neuroparalytic disease caused by a potent toxin produced by the spore-producing Gram-positive anaerobic bacterium, Clostridium botulinum. Mouse bioassays are a common standard assay for confirmation of botulism, but the assay is causing ethical concerns due to the use of mice and too slow for diagnosis. Therefore, fast and more reliable new detection methods are necessary. This study introduces sandwich ELISA kits for rapid detection of botulinum toxin A or B during bioterrorism or outbreaks. To verify the kits, sensitivity, precision, and specificity tests were conducted using human serum spiked with the toxins. The results showed limit of detections of 0.153 ng/ml and 0.027 ng/ml for toxins A and B, respectively, demonstrating strong repeatability and reproducibility within and between kits and experimenters (coefficient of variation: 10%). Specificity experiments confirmed accurate detection of only A and B toxins, without reacting to other toxins. The study establishes that the botulinum toxin A or B sandwich ELISA kits possess high sensitivity and specificity, effectively detecting toxin of levels comparable to the standard assay. These results suggest that the ELISA would serve as an effective test to detect botulinum toxin in the clinical specimens from the suspected patients. These findings suggest their potential in swift responses to biological terrorism and outbreaks.
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KEYWORD
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Botulism, Botulinum toxin, Sandwich ELISA, Mouse bioassay
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