Metcalf J S, Hyenstrand P, Beattie K A, Codd G A
Department of Biological Sciences, University of Dundee, UK.
J Appl Microbiol. 2000 Sep;89(3):532-8. doi: 10.1046/j.1365-2672.2000.01141.x.
Two types of commercially available ELISA kits for the immunoassay of cyanobacterial microcystins were evaluated for potential interference effects due to methanol, salinity, pH, plasticware and cyanobacterial extract. Of the treatments examined, methanol had the greatest effect, giving false positive microcystin concentrations with increasing methanol concentrations up to 30% (v/v) compared with the negative calibrators of each kit. False positive microcystin results were also produced with increasing salinity up to full strength seawater. Decreases in microcystin-LR equivalents were observed when assaying purified microcystin-LR at pH values between 6.25 and 10. Aqueous microcystin-LR solutions in plastic microcentrifuge tubes after pipetting with disposable plastic tips had lower toxin concentrations than expected when analysed by ELISA. Indicated microcystin concentrations in cyanobacterial extracts varied between kit types and the choice of blanks used. Although ELISAs can be useful tools for the screening of water and cyanobacterial blooms for microcystins and nodularins, users should be aware that commercial kits can be susceptible to interference by commonly encountered environmental and laboratory conditions and materials.
对两种市售的用于免疫测定蓝藻微囊藻毒素的ELISA试剂盒,评估了甲醇、盐度、pH值、塑料制品和蓝藻提取物可能产生的干扰效应。在所检测的处理中,甲醇的影响最大,与各试剂盒的阴性校准品相比,随着甲醇浓度增加至30%(v/v),会产生微囊藻毒素浓度的假阳性结果。盐度增加至全强度海水时也会产生微囊藻毒素的假阳性结果。在pH值介于6.25至10之间测定纯化的微囊藻毒素-LR时,观察到微囊藻毒素-LR当量降低。用一次性塑料吸头移液后,塑料微量离心管中的微囊藻毒素-LR水溶液经ELISA分析时,毒素浓度低于预期。蓝藻提取物中指示的微囊藻毒素浓度因试剂盒类型和所用空白的选择而异。虽然ELISA可作为筛选水体和蓝藻水华中微囊藻毒素和节球藻毒素的有用工具,但用户应意识到,市售试剂盒可能易受常见环境和实验室条件及材料的干扰。