Enzyme-liked immunosorbent assay for the detection of pathogenic spiroplasma in commercially exploited crustaceans from China
Enzyme-liked immunosorbent assay for the detection of pathogenic spiroplasma in commercially exploited crustaceans from China
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酶样免疫吸附试验用于检测中国商业化甲壳类动物中的致病性螺原体
DOI：
10.1016/j.aquaculture.2009.04.022
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发表时间：
2009-07
期刊：
影响因子：
4.5
通讯作者：
H. Huang
中科院分区：
文献类型：
--
作者：
K. Bi;JD. Shields;J. Wang;Q. Feng;W. Gu;T. Liang;W. Wang;H. Huang
螺旋体是在中国商业开发的淡水甲壳类动物中出现的新病原体。该细菌在中国江苏和浙江感染中华绒螯蟹（Eriocheir sinensis）、克氏原螯虾（Procambarus clarkii）和太平洋白虾（Litopenaeus vannamei）。本文制备了一种兔抗血清，用于对抗具有明显震颤病体征的中华绒螯蟹分离的螺旋体。建立了一种快速检测病原菌的间接酶联免疫吸附试验（ELISA）。抗体滴度为1:31,250。纯培养最低抗原蛋白检测值为0.573 μg/mL （3×105CCU·mL - 1pathogens）。与其他7种细菌（不动杆菌、枯草芽孢杆菌、链霉菌、铜绿假单胞菌、大肠杆菌、霍乱弧菌、嗜水气单胞菌）均无显色反应。在对植物和蜜蜂的螺原体进行的试验中观察到非常弱的显色反应。该检测的变异系数小于10%，在中度和重度感染中具有良好的可靠性。整个检测过程快速，可在3 h内完成。结果表明，该方法是诊断甲壳类螺原体疾病的一种简便易行的方法。
Spiroplasmas are emerging pathogens in commercially exploited freshwater crustaceans from China. The bacteria infect Chinese mitten crab (Eriocheir sinensis), crayfish (Procambarus clarkii) and pacific white shrimp (Litopenaeus vannamei) in Jiangsu and Zhejiang provinces of China. A rabbit antiserum was prepared against the spiroplasma isolated from a Chinese mitten crab that had obvious signs of tremor disease. An indirect enzyme-linked immunosorbent assay (ELISA) for the rapid detection of pathogen was developed. The titer of antibody against the pathogen was 1:31,250. The lowest detectable antigen protein was 0.573 μg/mL (3×105CCU·ml−1pathogens) in pure culture. No color reaction was observed with seven other bacteria (Acinetobacter spp, Bacillus subtilis, Streptomyces sp., Pseudomonas aeruginosa, Escherichia coli, Vibrio cholerae, Aeromonas hydrophila). Very weak color reaction was observed in tests with spiroplasma species from plants and honeybees. The coefficient of variability of the assay was less than 10%, with an excellent reliability in moderate and heavy infections. The whole test procedure was rapid, being completed within 3 h. The results showed that the assay is a simple and applied method for diagnosis of spiroplasma diseases in crustaceans.
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影响因子：
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作者：
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通讯作者：
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作者：
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