Chung D H, Abouzied M M, Pestka J J
Department of Food Science and Human Nutrition, Michigan State University, East Lansing 48824.
Mycopathologia. 1989 Sep;107(2-3):93-100. doi: 10.1007/BF00707544.
Conventional thin layer and instrumental methods for analyzing mycotoxins and their precursors are time-consuming and make the investigation of mycotoxin biosynthesis particularly difficult. As an alternative, sensitive enzyme-liked immunosorbent assays (ELISAs) can be utilized to analyze for these compounds. In this report, sterigmatocystin production in test tube cultures of Aspergillus versicolor ATCC 18643 and Aspergillus nidulans ATCC 32610 were compared using competitive ELISA. Polyclonal antiserum that was prepared against a sterigmatocystin hemiacetal-bovine serum albumin conjugate exhibited greatest specificity for sterigmatocystin hemiacetal and sterigmatocystin with less reactivity for O-methylsterigmatocystin. The antiserum could be used to detect as little as 50 ng/ml sterigmatocystin in ELISA. Direct ELISA could be performed on diluted culture broth and on mycelial extracts solubilized with N,N-dimethylformamide. Aspergillus versicolor ATCC 18643 produced more sterigmatocystin in SLS medium than in YES medium, and showed maximal levels at between 9 to 12 days incubation. Approximately 75% of sterigmatocystin was detectable in mycelium (254 micrograms/ml culture) compared to the extracellular fraction (87 micrograms/ml culture). Aspergillus nidulans exhibited qualitatively similar patterns of growth and toxigenesis in SLS medium but accumulated maximal levels of only 15 micrograms mycelial sterigmatocystin/ml culture and 5 micrograms extracellular sterigmatocystin/ml broth, respectively.
用于分析霉菌毒素及其前体的传统薄层法和仪器法耗时较长，使得霉菌毒素生物合成的研究尤为困难。作为一种替代方法，可以利用灵敏的酶联免疫吸附测定法（ELISA）来分析这些化合物。在本报告中，使用竞争性ELISA比较了杂色曲霉ATCC 18643和构巢曲霉ATCC 32610试管培养物中柄曲霉素的产生情况。针对柄曲霉素半缩醛 - 牛血清白蛋白偶联物制备的多克隆抗血清对柄曲霉素半缩醛和柄曲霉素表现出最大的特异性，而对O - 甲基柄曲霉素的反应性较低。该抗血清可用于在ELISA中检测低至50 ng/ml的柄曲霉素。直接ELISA可在稀释的培养液以及用N，N - 二甲基甲酰胺溶解的菌丝体提取物上进行。杂色曲霉ATCC 18643在SLS培养基中产生的柄曲霉素比在YES培养基中更多，并且在培养9至12天之间显示出最高水平。与细胞外部分（87微克/毫升培养液）相比，约75%的柄曲霉素可在菌丝体中检测到（254微克/毫升培养液）。构巢曲霉在SLS培养基中表现出质量上相似的生长和产毒模式，但分别仅积累了最高水平为15微克菌丝体柄曲霉素/毫升培养液和5微克细胞外柄曲霉素/毫升培养液。
