奶制品中三聚氰胺快速检测ELISA试剂盒

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1 Tribo TM Type 2 Shiga Toxin (Stx2) Test Strip User Guide (Catalog No. TBS11151) TribioScience, Inc.

2 General Description Enterohemorrhagic E. coli (EHEC) has become a world-wide epidemic, and was found in countries like US, Canada, Japan, China, and Germany, where a large outbreak of highly virulent E. coli O104:H4 occurred recently. There are multiple virulent factors in EHEC, with the Shiga Toxins being the most prominent. Among these, Type 2 Shiga Toxin (Stx2) carried the highest toxicity, which could lead to life threating Hemolytic Uremic Syndrome (HUS) in patients. Detection of Stx2 from EHEC greatly facilitates determination of strain virulence, and thereby improves the detection rate for highly virulent E. coli O104:H4 strain. In other words, when the sample tests positive on the test strip, it contains EHEC capable of producing Stx2 toxin, and therefore possesses high risk level. Tribo TM Type 2 Shiga Toxin (Stx2) test strip provides a convenient and rapid test for Stx2 toxin with a colloidal gold immunochromatographic design. It detects Stx2 with two different monoclonal antibodies in a sandwich assay format, and can be easily performed on feces, food, and water samples. It is a fast (results shown in 5-10 minutes), simple, sensitive and reliable approach for Stx2 detection. Intended Use Tribo TM Type 2 Shiga Toxin (Stx2) test strip is a lateral flow strip test for the qualitative detection of Stx2 produced by enterohemorrhagic E. coli (EHEC) in feces, food, and water samples. Safety Instructions To receive complete safety information on this product, please contact TribioScience, Inc. and request Material Safety Data Sheet. Assay Principles Tribo TM Stx2 test strip is based on sandwich immunoassay principle. An anti-ehec Stx2 monoclonal antibody is conjugated to colloidal gold and placed on conjugate pad. Colloidal gold provides red color to visualize antibody-antigen binding. Another anti-stx2 antibody is immobilized on nitrocellulose membrane. Test sample is loaded onto sample pad, and subsequently mixed with gold-antibody conjugate before migrating along the membrane. If the sample contains no Stx2, antibody conjugated to colloidal gold will not bind Stx2, and therefore will not bind the antibody immobilized on membrane. No red color will be observed on membrane detection line where the latter antibody is immobilized - 2 -

3 (indicating a negative result). If Stx2 is present in the test sample, it will bind gold-antibody conjugate and subsequently the antibody immobilized on membrane to form antibody-stx2-antibody sandwich-like complex. This complex will present a red colored band on detection line on membrane (as a positive result). A secondary anti-mouse IgG antibody is immobilized on membrane on a separate line (control line) to ensure the validity of each strip test. Reagents and Materials in each pack a) 1 Stx2 test strip b) 1 disposable dropper c) 1 pack of desiccant Sample Collection and Processing Optimal test results are obtained after amplification of bacteria from samples 1x bacteria growth media: modified EC broth plus 20 mg/l novobiocin For feces samples: Inoculate 1 g sample into 9 ml bacterial growth media, and shake for 6 h in 37 o C shaker-incubator For food samples: Grind 25 g sample and immediately inoculate into 225 ml bacterial growth media, and shake for 9 h in 37 o C shaker-incubator For water samples: Inoculate 400 ml sample into 100 ml 5x bacterial growth media, and shake for h in 37 o C shaker-incubator Test Procedure Equilibrate test strip at room temperature (20-25 o C) Equilibrate bacterial culture mentioned above to room temperature (20-25 o C) a) Remove test strip from package and place it on a dry, clean bench. b) Load 3 drops of cooled bacterial culture into sample loading well (S) and start timer. c) Observe result after 2-15 minutes. Results are invalid when observed beyond 15 minutes - 3 -

4 Result Interpretation Test result is interpreted by observing test line and control line shown in result window. Positive: both test line and control line present Negative: no test line, only control line present Invalid test: no control line present. Please repeat the test using a new test strip following instructions on this user guide. Positive Negative Invalid Sample Loading Sample Loading Sample Loading Test Line Test Line Test Line Control Line Control Line Control Line Precautions a) Test strip should be stored at room temperature and avoid freezing conditions b) Test strip is for one-time use c) After opening the package, use the strip on the same day d) Test is performed at room temperature, and test strip should be equilibrated to room temperature before use e) Test sample (bacterial culture after amplification) should be equilibrated to room temperature before being loaded onto strip Storage and Expiration Date Storage: keep at room temperature in a cool, dry place. Expiration Date: 12 months after manufacturing date. Technical Assistance - 4 -

5 For ordering or technical assistance regarding this product, or for additional information about TribioScience products, please or call (650) General Limited Warranty TribioScience, Inc. warrants its manufactured products against defects in materials and workmanship when used in accordance with the applicable instructions for a period not to extend beyond a product s printed expiration date. TribioScience makes no other warranty, expressed or implied. There is no warranty of merchantability or fitness for a particular purpose. The warranty provided herein and the data, specifications and descriptions of TribioScience products appearing in published catalogues and product literature may not be altered except by express written agreement signed by an officer of TribioScience. Representations, oral or written, which are inconsistent with this warranty or such publications are not authorized and, if given, should not be relied upon. In the event of a breach of the foregoing warranty, TribioScience Inc. s sole obligation shall be to repair or replace, at its option, any product or part thereof that proves defective in materials or workmanship within the warranty period, provided the customer notifies AccuAffinity promptly of any such defect. The exclusive remedy provided herein shall not be deemed to have failed of its essential purpose so long as TribioScience is willing and able to repair or replace any nonconforming TribioScience product or part. TribioScience shall not be liable for consequential, incidental, special or any other indirect damages resulting from economic loss or property damage sustained by a customer from the use of its products. However, in some states the purchaser may have rights under state law in addition to those provided by this warranty. For research use only

6 Tribo TM II 型志贺毒素 (Stx2) 快速检测试纸卡使用说明书 ( 产品编号 :TBS11151) 1. 产品概述 : 肠出血性大肠杆菌 (EHEC) 曾在世界各地流行, 包括美国加拿大日本中国, 以及最近暴发高致病性大肠杆菌 O104:H4 疫情的德国等 EHEC 含有多个毒性因子, 尤其是志贺毒素 (Shiga Toxin, Stx), 而其中又以 II 型志贺霉素 (Stx2) 的毒性最强, 能引起病人的溶血性尿毒综合征 (Hemolytic Uremic Syndrome, HUS), 进而危及生命 EHEC Stx2 的检测直接有助于判定分离菌株致病力的强弱, 从而提高对高致病性大肠杆菌 O104:H4 菌株的检出率若样品在该检测卡上显示阳性, 说明该样品中含有能产生 Stx2 毒素的 EHEC, 具有较高的风险等级本产品 II 型志贺毒素 (Stx2) 快速检测试纸卡将胶体金免疫层析原理应用于 II 型志贺毒素 (Stx2) 的快速测定, 对病毒抗原进行双抗体夹心检测用于快速筛查粪便, 食品, 水等样品中的肠出血性大肠杆菌 EHEC O104:H4 产生的 II 型志贺毒素 (Stx2), 协助食品安全检测与流行病学监控本产品使用简便, 反应灵敏检测迅速, 能够在 5-10 分钟内显示准确结果 2. 检测原理 : II 型志贺毒素 (Stx2) 快速检测试纸卡利用胶体金免疫层析技术原理, 将一种肠出血性大肠杆菌 Stx2 毒素 (EHEC Stx2) 的单克隆抗体包被于 NC 膜表面作为检测线 (T), 再将另一种 EHEC Stx2 单抗与胶体金偶联并附着在胶体金垫上当样品液体层析经过胶体金垫时, 样品中的 EHEC Stx2 毒素会与红色的单抗 - 胶体金偶联物特异性结合, 并继续层析至 NC 膜上与检测线位置包被的另一株单抗特异性结合, 形成一条粉红色带, 即显示出阳性结果当样品中不含有 EHEC Stx2 毒素时, 两株单抗无法在检测线位置形成双抗体夹心结构并显示粉红色条带, 这时检测结果显示为阴性 3. 试纸卡产品组成 : 试纸卡 (1 块 ); 一次性样品收集滴管 (1 支 ); 干燥剂 (1 块 ) 4. 样本收集及处理 : 样本增菌后效果检测较好 1 倍细菌培养液成份 : 改良 EC 肉汤加 20mg/L 新生霉素粪便样本 : 取 1g 样本, 立即接种于 9ml 的细菌培养液中,37 恒温摇床培养 6h - 6 -

7 食物样本 : 取 25g 样本, 磨碎, 立即接种于 225ml 的细菌培养液中,37 恒温摇床培养 9h 水源样本 : 取 400ml 样本, 立即接种于 100ml 的 5 倍浓缩细菌培养液中,37 恒温摇床培养 16-20h 5. 检测步骤 : (1) 检测卡放置于洁净干燥的工作台上 ; (2) 用滴管加三滴冷却后的增菌培养液到检测卡的加样孔 (S), 开始计时 ; (3) 2-15 分钟内观察结果,15 分钟之后判读结果无效 6. 结果判定 : 阴性 (-): 检测线 (T) 无色, 控制线 (C) 显色阳性 (+): 检测线 (T) 和控制线 (C) 都显色无效 : 控制线 (C) 无色, 表示使用方法不正确或试纸卡失效, 结果无效请对照使用说明书, 用新的试纸卡重新测定阴性阳性检测无效加样孔加样孔加样孔 T 检测线 T 检测线 T 检测线 C 控制线 C 控制线 C 控制线 7. 注意事项 : (1) 本产品试纸卡为一次性使用打开包装后, 请在当日尽快使用 (2) 检测过程请佩戴乳胶手套和口罩, 检测后请妥善处理具有潜在传染性的废弃物 8. 储存条件和有效期 : II 型志贺毒素 (Stx2) 快速检测试纸卡原包装在室温 (20-25 o C) 下储存, 置于阴凉避光干燥处有效期为 12 个月, 产品批号及有效期见包装袋本产品仅供研究使用 - 7 -

奶制品中三聚氰胺快速检测ELISA试剂盒

奶制品中三聚氰胺快速检测ELISA试剂盒 Tribo TM Aflatoxin B1 Rapid Test Strip User Guide (Catalog No. TBS11166) TribioScience, Inc. General Description Aflatoxins are a class of structurally similar mycotoxins produced by Aspergillus species,