斑点叉尾鮰（Ictalurus punctatus）肌肉中磺胺二甲氧嘧啶残留的提取与酶免疫测定

Extraction and enzyme immunoassay of sulfadimethoxine residues in channel catfish (Ictalurus punctatus) muscle.

作者信息

Walker C C, Barker S A

机构信息

Louisiana State University, School of Veterinary Medicine, Department of Veterinary Physiology, Pharmacology, and Toxicology, Baton Rouge 70803-8420.

出版信息

J AOAC Int. 1994 Jul-Aug;77(4):908-16.

PMID:8069122

Abstract

Four commercially available enzyme immunoassays were evaluated for the detection of sulfadimethoxine (SDM) residues in channel catfish muscle. Matrix solid-phase dispersion (MSPD) extracts of samples (n = 60, 0.5 g) fortified with SDM at 0, 25, 50, 100, and 250 ng/g were used in all assays. Intra-assay variations for 2 MICROTITER well-based quantitative assays, SIGNAL sulfamethazine test and IDS SDM One-Step ELISA (enzyme-linked immunosorbent assay), were 5.6 and 7.7%. Interassay variations for these tests were 7.9 and 16.6%. The agreements between evaluators for 2 membrane-based, visually determined assays, CITE Sulfa Trio and EZ-SCREEN: SDM, were 77 and 95%. Performance values, as indicated by sensitivity, specificity, efficiency, and positive and negative predictive values, were 100, 92, 95, 89, and 100%, respectively for the SIGNAL test; 100, 94, 97, 92, and 100% for the IDS test; 98, 71, 82, 69, and 98% for the CITE test; and 98, 94, 96, 92, and 99% for the EZ-SCREEN assay. Eight sulfonamides cross-reacted in the SIGNAL test; EC-50 values (concentrations causing 50% inhibition of color development compared with blanks) varied from < 0.1 to 45 micrograms/mL. The EC-50 value for SDM was 0.25 microgram/mL. The CITE test cross-reacted with sulfachloropyridazine at 10 micrograms/mL. The IDS and EZ-SCREEN tests had no significant cross-reactivity with other sulfonamides. N-Acetyl SDM reacted like the parent SDM in all assays. Performance results indicated that MSPD extracts of catfish muscle may be used in these immunoassays to screen catfish muscle samples for violative levels of SDM residue.

摘要

对四种市售酶免疫测定法进行了评估，以检测斑点叉尾鮰肌肉中的磺胺二甲氧嘧啶（SDM）残留。在所有测定中均使用了用0、25、50、100和250 ng/g的SDM强化的样品（n = 60，0.5 g）的基质固相分散（MSPD）提取物。两种基于微孔板的定量测定法，即SIGNAL磺胺二甲嘧啶试验和IDS SDM一步酶联免疫吸附测定法（ELISA）的批内变异分别为5.6%和7.7%。这些试验的批间变异分别为7.9%和16.6%。两种基于膜的目视测定法，即CITE Sulfa Trio和EZ-SCREEN：SDM，评估者之间的一致性分别为77%和95%。SIGNAL试验的性能值，以灵敏度、特异性、效率以及阳性和阴性预测值表示，分别为100%、92%、95%、89%和100%；IDS试验为100%、94%、97%、92%和100%；CITE试验为98%、71%、82%、69%和98%；EZ-SCREEN测定法为98%、94%、96%、92%和99%。八种磺胺类药物在SIGNAL试验中发生交叉反应；半数有效浓度（EC-50值，即与空白相比导致显色抑制50%的浓度）在<0.1至45微克/毫升之间变化。SDM的EC-50值为0.25微克/毫升。CITE试验与10微克/毫升的磺胺氯哒嗪发生交叉反应。IDS和EZ-SCREEN试验与其他磺胺类药物无显著交叉反应。N-乙酰基SDM在所有测定中的反应与母体SDM相似。性能结果表明，斑点叉尾鮰肌肉的MSPD提取物可用于这些免疫测定法，以筛选斑点叉尾鮰肌肉样品中是否存在违规水平的SDM残留。