以直接酵素連結免疫吸附分析試劑測定豬體中磺胺二甲嘧啶   全文下載

Deter ruination of Sulfamethazine in Swine Serum with One-step ELISA Test Kit

磺胺二甲嘧啶（sulfamethazine）被懷疑具有致癌性，在動物飼料添加劑使用非常廣泛。為減少豬肉磺胺二甲嘧啶高殘留率，必須實施全國屠宰場的監視作業。傳統的檢驗方法耗時費事，因而有磺胺二甲嘧啶直接酵素連結免疫吸附劑分析法產生以作快速篩選檢測。本研究即在評估市售美國國際診斷系統公司生產之磺胺二甲嘧啶直接酵素連結免疫吸附分析試劑套組（sulfamethazine one-step enzyme-linked immunosorbent assay test kit）之性能，其結果可做為使用此試劑者之參考。包括樣品前處理，該試劑所需檢測時間約30分鐘，在平板振盪器上一次可同時分析96個樣品，其偵測極接度（detection limit）為0.001 ppm。於空白的血清樣品添加0.04、0.2和0.42 μg／ml，經六重複試驗結果，平均回收率為99.8%（ 變異係數2.92%）。以11種常用磺胺劑：sulfamethazine, sulfamerazine, sulfadimethoxine, sulfamonomethoxine, sulfathiazole, sulfamethoxypyridazine, sulfaquinoxaline, sulfamethoxazole, sulfapyridine, sulfisoxazole和sulfadiazine 各以四種不同濃度（包括0.001、0.01、0.1和1.0 μg／ml），進行試劑套組中之交叉反應性測試。若設定50%為抑制呈色反應干擾成立值，則上述11種磺胺劑濃度在1.0 μg／ml時僅有sulfamerazine、sulfisoxazole和sulfapyridine等三種磺胺劑有干擾現象。若以sulfamethazine之相對反應性為100%計算，則sulfamerazine為10.0%，sulfisoxazole為3 .4%，sulfpyridine為2.0%。當上述三種磺胺劑濃度降低至0.1 μg／ml時則不受交叉反應性之影響。此試劑組操作簡便、敏感度高、檢測時間短、安全無污染且檢測成本日漸低廉，故可供作為豬肉中磺胺二甲嘧啶殘留之快速定性及半定量篩檢用。"

Sulfamethazine is a suspected carcinogen. It is widely used as feed additive for swine rations. An effective way of reducing this high incidence of sulfamethazine violations in swine would be to implement a nationwide slaughterhouse surveillance program for the drug. The traditional method for the detection of sulfamethazine spends too much time. A rapid screening test of sulfamethazine residue in swine serum was made by the use of the sulfamethazine one-stop Enzyme- Linked Immunosorbent Assay (ELISA) Kit. The time for sample preparation and reaction in microwells and read in ultrascan meter was about 30 minutes. The detection limit of sulfamethazine was 0.00 I ppm. The mean recoveries of added sulfamethazine from serum at levets 0.04, 0.2, and 0.42 µg/ml were 99.8% (coefficient of variation, CV=2.92%). Among 11 sulfonamide analogus: sulfamethazine, sulfamerazine, sulfadimethoxine, sulfamonomethoxine, sulfathiazole, sulfamethoxypyridazine, sulfaquinoxaline, sulfamethoxazole, sulfapyridine, sulfisoxazole and sulfadiazine with 4 different concentrations : 0.001, 0.01, 0.1 and 1.0 µg/ml had cross reaction to antibodies in the ELISA Kit. The relative reactivity of sulfamethazine, sulfamerazine, sulfisoxazole and sulfapyridine were 100, 10.0, 3.4 and 2.0%, respectively. The concentration of sulfonamide below 0.1 µg/ml, the cross reaction was neglected. This ELISA kit may be used as a rapid screening test for sulfamethazine residue in swine serum.