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中华鳖全血呼吸爆发化学发光检测方法的建立
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张敏， 李肖梁，邹晓庭
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（浙江大学 动物科学学院， 浙江 杭州310029）
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摘要：
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为进一步研究中华鳖的先天免疫能力及其机理，以酵母多糖为吞噬底物，通过对鲁米诺浓度、全血稀释度和检测温度等条件的优化，建立了中华鳖全血呼吸爆发的化学发光检测方法，并用此方法对放置不同时间的全血和稀释血液进行了呼吸爆发检测效果的比较。结果表明，鲁米诺浓度为1×10-4 mol/L，检测温度为315℃时，以04~08μL/mL的中华鳖全血进行检测可获得理想效果。保存6d的全血仍可检测到呼吸爆发，稀释全血放置30h后不再产生呼吸爆发。 |
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关键词：
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中华鳖； 呼吸爆发； 全血； 化学发光 |
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中图分类号：S 858.99： Q 47 文献标识码：
A 文章编号：1673-4696(2006)02-0139-04 |
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Development of chemoluminescence method for detection of respiratory
burst activity of Trionyx sinensis whole blood |
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ZHANG Min, LI Xiaoliang, ZOU Xiaoting |
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(College of Animal Science, Zhejiang University, Hangzhou 310029, China) |
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Abstract:The system of Trionyx sinensis whole blood respiratory burst activity experimentation was explored by optimizing luminols′s concentration, the whole blood dilution of turtle and temperature when zymosan was used as the substrata of the phagocytosis. The detected results were good at 1×10-4 mol/L of luminol′s concentration, 315 ℃ and 04~08μL/mL of turtles′ whole blood. The respiratory burst activity was still detectable using the turtle’s whole blood reserved for 6 d, however the peak value of respiratory burst could not be detected in the diluted turtles’ whole blood which had been diluted and detained for 30 h. The development of chemoluminescence method for the detection of respiratory burst activity in Trionyx sinensis whole blood provided a novel alternative means of further exploring on innate immunity of Trionyx sinensis. |
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Key words: Trionyx sinensis; respiratory burst； whole blood; chemoluminescence |
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