A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid.
A competitive ELISA to detect brevetoxins from Karenia brevis (formerly Gymnodinium breve) in seawater, shellfish, and mammalian body fluid.
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一种竞争性ELISA，可检测海水，贝类和哺乳动物体液中的Karenia Brevis（以前是Bredinium Breve）的Brevetoxins。
DOI：
10.1289/ehp.02110179
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发表时间：
2002-02
影响因子：
10.4
通讯作者：
Baden DG
中科院分区：
文献类型：
--
作者：
Naar J;Bourdelais A;Tomas C;Kubanek J;Whitney PL;Flewelling L;Steidinger K;Lancaster J;Baden DG
我们开发了一种竞争性的酶联免疫吸附试验（ELISA）分析brevetoxins，使用山羊抗brevetoxin抗体免疫后获得的匙孔血蓝蛋白-brevetoxin共轭物，结合三步信号放大过程。该程序，其中使用二级生物素化抗体，链霉亲和素-辣根过氧化物酶结合物，和显色酶底物，是有用的，在减少非特异性背景信号通常观察到的复杂的矩阵。这种竞争性ELISA检测海水，贝类提取物和匀浆，和哺乳动物体液，如尿液和血清中的短尾藻毒素未经预处理，稀释，或纯化。我们调查了应用这种技术的贝类监测，通过加标贝类肉与brevetoxins和分析牡蛎从两个商业贝类床在佛罗里达暴露于开花的Karenia brevis（原Gymatriinium breve）。我们进行了贝类提取物和匀浆中的短尾轮虫毒素的ELISA分析，并与小鼠生物测定法和受体结合试验进行了比较。在加标的牡蛎中，短尾藻毒素的检测限为2.5微克/100克贝类肉。这种测定似乎是一个有用的工具，贝类和海水中的神经毒性贝类中毒监测，哺乳动物暴露诊断，并显着减少所需的分析时间。
We developed a competitive enzyme-linked immunosorbent assay (ELISA) to analyze brevetoxins, using goat anti-brevetoxin antibodies obtained after immunization with keyhole limpet hemocyanin-brevetoxin conjugates, in combination with a three-step signal amplification process. The procedure, which used secondary biotinylated antibodies, streptavidine-horseradish peroxidase conjugate, and chromogenic enzyme substrate, was useful in reducing nonspecific background signals commonly observed with complex matrices. This competitive ELISA detected brevetoxins in seawater, shellfish extract and homogenate, and mammalian body fluid such as urine and serum without pretreatment, dilution, or purification. We investigated the application of this technique for shellfish monitoring by spiking shellfish meat with brevetoxins and by analyzing oysters from two commercial shellfish beds in Florida that were exposed to a bloom of Karenia brevis (formerly Gymnodinium breve). We performed brevetoxin analysis of shellfish extracts and homogenates by ELISA and compared it with the mouse bioassay and receptor binding assay. The detection limit for brevetoxins in spiked oysters was 2.5 microg/100 g shellfish meat. This assay appears to be a useful tool for neurotoxic shellfish poisoning monitoring in shellfish and seawater, and for mammalian exposure diagnostics, and significantly reduces the time required for analyses.
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DOI：
10.1080/009841099157656
发表时间：
1999-07-09
期刊：
JOURNAL OF TOXICOLOGY AND ENVIRONMENTAL HEALTH-PART A
影响因子：
--
作者：
Benson, JM;Tischler, DL;Baden, DG
通讯作者：
Baden, DG
影响因子：
4.7
作者：
Naar, J;Branaa, P;Pauillac, S
通讯作者：
Pauillac, S
影响因子：
2.4
作者：
GERACI, JR;ANDERSON, DM;MAYO, CA
通讯作者：
MAYO, CA
影响因子：
2.8
作者：
BADEN, DG;MENDE, TJ;SCHULTZ, DR
通讯作者：
SCHULTZ, DR
影响因子：
2.8
作者：
LEVINE, L;SHIMIZU, Y
通讯作者：
SHIMIZU, Y