人（Human）腺病毒IgM抗体（ADV-IgM）-定性A

人（Human）腺病毒IgM抗体（ADV-IgM）-定性A 2. 血浆,EDTA、柠檬酸盐或肝素抗凝。3000转离心30分钟取上清。 本试剂盒只能用于科学研究,不得用于医学诊断 3. 细胞上清液,3000转离心10分钟去除颗粒和聚合物。 人，Human，腺病毒IgM抗体，ADV-IgM， 4. 组织匀浆,将组织加入适量生理盐水捣碎。3000转离心10分钟ELISA检测试剂盒 使用说明书 取上清。 检测原理 5. 保存,如果样本收集后不及时检测,请按一次用量分装,冻存于 试剂盒采用双抗一步夹心法酶联免疫吸附试验，ELISA，。往预先-20?,避免反复冻融,在室温下解冻并确保样品均匀地充分解冻。 包被抗原的包被微孔中,依次加入标本、HRP标记的检测抗原,经过自备物品 温育并彻底洗涤。用底物TMB显色,TMB在过氧化物酶的催化下转化1. 酶标仪，450nm， 成蓝色,并在酸的作用下转化成最终的黄色。用酶标仪在450nm波长2. 高精度加样器及枪头,0.5-10uL、2-20uL、20-200uL、200-1000uL 下测定吸光度，OD 值，,与CUT OFF值相比较,从而判定标本中人腺3. 37?恒温箱 病毒IgM抗体，ADV-IgM，的存在与否。 操作注意事项 样品收集、处理及保存 1. 试剂盒保存在2-8?,使用前室温平衡20分钟。从冰箱取出的1. 血清,使用不含热原和内毒素的试管,操作过程中避免任何细胞浓缩洗涤液会有结晶,这属于正常现象,水浴加热使结晶完全溶解刺激,收集血液后,3000转离心10分钟将血清和红细胞迅速小心地后再使用。 分离。 2. 实验中不用的板条应立即放回自封袋中,密封，低温干燥，保存。 3. 预处理后的样本无需稀释,直接取50μL加样即可。 1. 手工洗板,甩尽孔内液体,每孔加满洗涤液,静置1min后甩尽4. 严格按照说明书中标明的时间、加液量及顺序进行温育操作。 孔内液体,在吸水纸上拍干,如此洗板5次。 5. 所有液体组分使用前充分摇匀。 2. 自动洗板机,每孔注入洗液350μL,浸泡1min,洗板5次。 试剂盒组成 操作步骤 名称 96孔配置 48孔配置 备注 1. 从室温平衡20min后的铝箔袋中取出所需板条,剩余板条用自封微孔酶标板 12孔×8条 12孔×4条 无 阴性对照 0.5mL 0.5mL 无 袋密封放回4?。 阳性对照 0.5mL 0.5mL 无 检测抗原-HRP 10mL 5mL 无 2. 设置阴、阳性对照孔和样本孔,阴、阳性对照孔中加入阴性对照、20×洗涤缓冲液 25mL 15mL 按说明书进行稀释 底物A 6mL 3mL 无 阳性对照各50μL, 底物B 6mL 3mL 无 终止液 6mL 3mL 无 3. 待测样本孔加待测样本50μL, 封板膜 2张 2张 无 说明书 1份 1份 无 4. 随后阴、阳性对照孔和样本孔中每孔加入辣根过氧化物酶，HRP，自封袋 1个 1个 无 标记的检测抗原100μL,用封板膜封住反应孔,37?水浴锅或恒温试剂的准备 箱温育60min。 20×洗涤缓冲液的稀释,蒸馏水按1,20稀释,即1份的20×洗涤缓5. 弃去液体,吸水纸上拍干,每孔加满洗涤液,静置1min,甩去冲液加19份的蒸馏水。 洗涤液,吸水纸上拍干,如此重复洗板5次，也可用洗板机洗板，。 洗板方法 6. 每孔加入底物A、B各50μL,37?避光孵育15min。 7. 每孔加入终止液50μL,15min内,在450nm波长处测定各孔的免责声明 OD值。 1. 试剂盒仅供研究使用,不得用于临床实验或人体实验,否则所结果判断 产生的一切后果,由实验者承担,本公司概不负责。 1. 试验有效性,阳性对照孔OD值平均值?1.00, 2. 严格按照说明书操作,实验者违反说明书操作,后果由实验者 阴性对照孔OD值平均值?0.15。 承担。 2. 临界值，Cut off，计算,临界值=阴性对照孔平均值+0.15 3. 阴性判断,样品OD值,临界值，Cut off，,样品为阴性 4. 阳性判断,样品OD值,临界值，Cut off，,样品为阳性 试剂盒性能 1. 准确性,阳性对照孔OD值平均值?1.00,阴性对照孔OD值平均 值?0.15,说明试验结果有效。 2. 特异性,不与其它可溶性结构类似物交叉反应。 3. 重复性,板内、板间变异系数均小于15%。 4. 贮藏,2-8?,避光防潮保存。 5. 有效期,6个月 FOR RESEARCH USE ONLY. assay immediately or aliquot and store samples at -20? or -80?.Avoid repeated NOT FOR USE IN DIAGNOSTIC PROCEDURES. freeze-thaw cycles Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge Human Adenovirus IgM (ADV-IgM) ELISA Kit samples for 30 minutes at 3000×g at 2-8? within 30 minutes of collection. Store samples at -20?or -80?. Avoid repeated freeze-thaw cycles. instruction Cell culture supernates and other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20?or Intended use -80?. Avoid repeated freeze-thaw cycles. This ADV-IgM ELISA kit is intended Laboratory for Research use only and is not Note: The samples shoule be centrifugated dequately and no hemolysis for use in diagnostic or therapeutic procedures.The Stop Solution changes the color or granule was allowed. from blue to yellow and the intensity of the color is measured at 450 nm using a Materials required but not supplied spectrophotometer. In order to measure the concentration of ADV-IgM in the sample, this ADV-IgM ELISA Kit includes a set of calibration standards. The 1. Standard microplate reader(450nm) calibration standards are assayed at the same time as the samples and allow the 2. Precision pipettes and Disposable pipette tips. operator to produce a standard curve of Optical Density versus ADV-IgM 3. 37 ? incubator concentration. The concentration of ADV-IgM in the samples is then determined by Precautions comparing the O.D. of the samples to the standard curve. 1. Do not substitute reagents from one kit to another. Standard, conjugate and Sample collection and storages microplates are matched for optimal performance. Use only the reagents supplied by manufacturer. Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 10 minutes at approximately 3000×g. Remove serum and 2. Do not remove microplate from the storage bag until needed. Unused strips should be stored at 2-8?C in their pouch with the desiccant provided. 2. Separately add Positive control and Negative control 50μl to the Positive and 3. Mix all reagents before using. Negative well, add 50μl of Sample to testing sample well. Remove all kit reagents from refrigerator and allow them to reach room temperature 3. Add 100μl of HRP-conjugate reagent to each well, cover with an adhesive strip ( 20-25?C) and incubate for 60 minutes at 37?C. 4. Aspirate each well and wash, repeating the process four times for a total of five Materials supplied washes. Wash by filling each well with Wash Solution (400μl) using a squirt bottle, Name 96 determinations 48 determinations Microelisa stripplate 12*8strips 12*4strips manifold dispenser or autowasher. Complete removal of liquid at each step is Negative control 0.5ml 0.5ml essential to good performance. After the last wash, remove any remaining Wash Positive control 0.5ml 0.5ml Solution by aspirating or decanting. Invert the plate and blot it against clean paper HRP-Conjugate reagent 10.0ml 5.0ml 20X Wash solution 25ml 15ml towels. Chromogen Solution A 6.0ml 3.0ml 5. Add chromogen solution A 50μl and chromogen solution B 50μl to each well. Chromogen Solution B 6.0ml 3.0ml Gently mix and incubate for 15 minutes at 37?C. Protect from light. Stop Solution 6.0ml 3.0ml 6. Add 50μl Stop Solution to each well. The color in the wells should change Closure plate membrane 2 2 User manual 1 1 from blue to yellow. If the color in the wells is green or the color change does not Sealed bags 1 1 appear uniform, gently tap the plate to ensure thorough mixing. Reagent preparation 7. Read the Optical Density (O.D.) at 450 nm using a microtiter plate reader 20×wash solution:Dilute with Distilled or deionized water 1:20. within 15 minutes. Assay procedure 1. Prepare all reagents before starting assay procedure. It is recommended that all Standards and Samples be added in duplicate to the Microelisa Stripplate. Determine the result 1. Test validity: the average of Positive control well?1.00; the average of Negative control well ?0.15. 2. Calculate Critical (CUT OFF): Critical= the average of Negative control well + 0.15. Negative Result: sample OD< Calculate Critical (CUT OFF) is Negative. Positive Result: sample OD? Calculate Critical (CUT OFF) is Positive. Storage and validity Storage: 2-8?. validity: six months. FOR RESEARCH USE ONLY; NOT FOR THERAPEUTIC OR DIAGNOSTIC APPLICATIONS! PLEASE READ THROUGH ENTIRE PROCEDURE BEFORE BEGINNING!