Rosselot G, Lopez-Lastra M, McMurtry J P
Instituto de Nutrición y Technología de los Alimentos, Universidad de Chile, Macul, Santiago, Chile.
Poult Sci. 1996 Jul;75(7):873-80. doi: 10.3382/ps.0750873.
A homologous RIA for the determination of toxic gizzerosine activity in commercial fish meals has been developed. Three polyclonal antibodies (GR316, GR415, and GR418) were produced in female rabbits and extensively characterized for their potential use in individual RIA. The RIA had lower detection limits of 0.048, 0.78, and 0.39 ng/mg using the three respective antibodies. Because gizzerosine is derived from lysine and histidine, crossreactivity with these amino acids, and with histamine was examined. The antibodies crossreacted with histamine from 21 to 100%. No crossreactivity with histidine or lysine was observed for any of the three antibodies. Antibody GR415 was chosen for determination of gizzerosine in extracted fish meal samples because crossreactivity of histamine using this antibody was only present at high concentrations, and the Ka value for gizzerosine was 10-fold greater than for histamine. A mild buffer extraction procedure was used, resulting in 98% gizzerosine recovery. Displacement curves from extracted and serially diluted fish meal samples were parallel with gizzerosine standard. Inter-and intra-assay coefficients of variation were 11 and 15%, respectively. We used the RIA for determination of gizzerosine activity in a pool of 23 fish meal samples of known gizzerosine scores (determined with a chick bioassay), and histamine content. The partial correlation coefficient between gizzerosine content determined by the RIA and gizzerosine scores from the bioassay was high (0.83), and significant (P < 0.01). There were also significant correlations between gizzerosine scores and histamine content of the fish meals (0.63, P < 0.01), and between histamine content and gizzerosine levels determined by the RIA (0.59, P < 0.01). The application of the homologous RIA for the determination of gizzerosine activity in commercial fish meals could be of importance for the prevention of gizzard erosions in the poultry industry, and for studying gizzerosine-induced pathology and metabolism.
已开发出一种用于测定商业鱼粉中毒性肌胃糜烂素活性的同源放射免疫分析方法。用雌性兔子制备了三种多克隆抗体（GR316、GR415和GR418），并对其在个体放射免疫分析中的潜在用途进行了广泛表征。使用这三种抗体时，该放射免疫分析的检测下限分别为0.048、0.78和0.39 ng/mg。由于肌胃糜烂素由赖氨酸和组氨酸衍生而来，因此检测了其与这些氨基酸以及组胺的交叉反应性。这些抗体与组胺的交叉反应率为21%至100%。三种抗体中任何一种与组氨酸或赖氨酸均未观察到交叉反应。选择抗体GR415用于测定提取的鱼粉样品中的肌胃糜烂素，因为使用该抗体时组胺的交叉反应仅在高浓度时出现，且肌胃糜烂素的Ka值比组胺大10倍。采用了温和的缓冲液提取程序，肌胃糜烂素的回收率为98%。提取并连续稀释的鱼粉样品的置换曲线与肌胃糜烂素标准曲线平行。批间和批内变异系数分别为11%和15%。我们使用该放射免疫分析方法测定了一组23个已知肌胃糜烂素评分（通过雏鸡生物测定法确定）和组胺含量的鱼粉样品中的肌胃糜烂素活性。放射免疫分析测定的肌胃糜烂素含量与生物测定法得到的肌胃糜烂素评分之间的偏相关系数很高（0.83），且具有显著性（P < 0.01）。鱼粉的肌胃糜烂素评分与组胺含量之间（0.63，P < 0.01），以及组胺含量与放射免疫分析测定的肌胃糜烂素水平之间（0.59，P < 0.01）也存在显著相关性。同源放射免疫分析方法用于测定商业鱼粉中的肌胃糜烂素活性，对于预防家禽业中的肌胃糜烂、研究肌胃糜烂素诱导的病理学和代谢可能具有重要意义。
