BioReagent 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
粪便的化学检查有酸碱度反应、隐血试验、胆色素和脂肪测定等，其中隐血试验最具有重要的临床应用价值，上消化道出血量小于 5ml，粪便中无可见的血液，且红细胞破坏，显微镜检查也未见红细胞，而需要用化学法、免疫法、转铁蛋白法、血红蛋白荧光测定法等才能证实的出血，称为隐血，检查粪便隐血的试验称为粪便隐血试验 (fecal occult blood test,FOBT)，化学法常用试剂有邻联甲苯胺、匹拉米洞、愈创木脂、无色孔雀绿、米土尔等。
本品的检测原理是血红蛋白中的亚铁血红素具有类似过氧化物酶的活性，能催化 H₂O₂作为电子受体使色原底物邻联甲苯胺氧化成邻甲偶氮苯而显绿色→蓝色，或使色原底物匹拉米洞氧化显紫色或紫蓝色，反应物颜色深浅与血红蛋白含量成正比，根据显色的快慢可以半定量血红蛋白即隐血含量，因此也可称作粪便隐血半定量试验，多用于人或动物粪便血红蛋白检查。应注意的是正常人或动物消化道也会微量出血或受到刺激后出血或模型动物饲料含有鱼粉、铁剂等，都可能导致试验结果出现假阳性，邻联甲苯胺法特点是灵敏度高、假阳性高，可检出 Hb 0.2~1.0mg/L，匹拉米洞法特点是灵敏度中、假阳性中，可检出 Hb 1~5mg/L，联合使用两种方法能更有效的判断隐血情况。该试剂盒仅用于科研领域，不适用于临床诊断或其他用途。
| 组分货号 | 组分名称 | 物理外观 | 2×100T | 2×300T | 储存 |
| F1527137A | 邻联甲苯胺溶液 | 液体 | 10ml | 30ml | 2-8℃.避光 |
| F1527137B | 匹拉米洞溶液 | 液体 | 10ml | 30ml | 2-8℃.避光 |
| F1527137C | 氧化剂 | 液体 | 2×10ml | 2×30ml | 2-8℃.避光 |
自备材料
1、 粪便标本、木片或牙签、消毒棉或白瓷板、滴管或移液器
操作步骤：(仅供参考)
1、粪便标本应尽快收集并及时检测，避免长时间放置使反应的灵敏度降低，避免接触水。
2、用取样木片或牙签挑取少量粪便 10~50mg（绿豆大小），涂在消毒棉或白瓷板上。
3、邻联甲苯胺法：滴加邻联甲苯胺溶液 2 滴(约 0.1ml)于粪便不同位置。匹拉米洞法：滴加匹拉米洞溶液 2 滴(约 0.1ml)于粪便不同位置。
4、滴加氧化剂 2 滴(约 0.1ml)，立即计时并观察颜色变化。
5、邻联甲苯胺法：于 2min 内判读完毕，如 2min 内有蓝色出现，提示粪便含有血红蛋白(Hb)或粪便隐血试验阳性。匹拉米洞法：于 2min 内判读完毕，如 2min 内有紫色出现，提示粪便含有血红蛋白(Hb)或粪便隐血试验阳性。
6、试剂开封后请尽快使用，以防影响后续实验效果。
结果判读
| 判读 | 颜色变化(邻联甲苯胺法) | 颜色变化(匹拉米洞法) |
|---|---|---|
| (4+) | 加入试剂后立即呈现蓝黑褐色 | 加入试剂后立即呈现深紫色 |
| (3+) | 加入试剂后立即呈现蓝褐色，并逐渐呈黑褐色 | 加入试剂后立即呈现紫色，并逐渐呈深紫色 |
| (2+) | 加入试剂后初显浅绿色，逐渐呈明显蓝褐色 | 加入试剂后初显浅紫色，逐渐呈明显紫色 |
| (+) | 10s 后由浅绿色渐变为绿色 | 10s 后由无色渐变为浅紫色或紫色 |
| 阴性 | 2min 内不显色 | 2min 内不显色 |
注意事项
1、在粪便标本采集、运送过程中和使用试剂进行检测时，操作人员应做好个人防护，避免人体接触，以防止可能产生的污染和自身感染。
2、正常人或动物消化道也会微量出血或受到刺激后出血，导致本法有时会出现假阳性。
3、对无任何明显症状但怀疑可能有少量出血的情况，建议至少连续 3 天取样，并从不同样本部位取材做两次实验。
4、应提前 3 天以上禁止食用动物血、肉、肝脏及富含叶绿素食物、铁剂、中药或其他特殊药物，以免出现假阳性反应。
5、对于结肠炎模型的实验动物，饲料成分不应含有鱼粉、铁剂或其他特殊药物，以免出现假阳性反应。
6、邻联甲苯胺溶液、匹拉米洞溶液含有乙酸，刺激性比较大，应在通风橱里操作，本溶液颜色可随配置时间的长短而发生变化，但不影响其检测的灵敏度。
7、氧化剂应注意密封 4℃保存，6 个月之后效率可能会下降，可考虑市售的医用过氧化氢或过氧化脲配置 1~3% 的溶液。
8、如有必要可用血液或血红蛋白溶液做梯度对照实验。
9、该试剂盒不能与氧化剂共同存放，并避免阳光直射及高低温储存环境。
10、为了您的安全和健康，请穿实验服并戴一次性手套操作。
有效期
12 个月有效；低温运输，4℃保存。
Chemical examinations of feces include pH reaction, fecal occult blood test, bile pigment assay and fat determination, among which the fecal occult blood test possesses high‑value clinical application. Occult bleeding refers to bleeding from the upper gastrointestinal tract with a volume less than 5 mL, where no visible blood is observed in feces, erythrocytes are degraded and undetectable under microscopy. Such bleeding can only be confirmed by chemical methods, immunoassays, transferrin‑based assays, hemoglobin fluorescence assays and other approaches. The assay for detecting fecal occult blood is defined as the fecal occult blood test (FOBT). Commonly‑used reagents for chemical methods include o‑tolidine, pyramidon, guaiac, leucomalachite green, metol, etc.
The detection principle of this kit: Heme in hemoglobin exhibits peroxidase‑like activity. It catalyzes H₂O₂ acting as an electron acceptor to oxidize the chromogenic substrate o‑tolidine into o‑azotoluene, producing a green‑to‑blue color; alternatively, it oxidizes the chromogenic substrate pyramidon to yield purple or bluish‑purple color. The color intensity of reaction products is proportional to hemoglobin concentration. Hemoglobin (i.e. occult blood content) can be semi‑quantified according to color‑development rate, hence this assay is also termed semi‑quantitative fecal occult blood test, which is mostly applied for hemoglobin detection in human or animal feces.
Note that trace gastrointestinal bleeding or irritation‑induced bleeding may occur in normal humans and animals; fish meal, iron supplements or other components in diet of model animals may also cause false‑positive results. The o‑tolidine method features high sensitivity and high false‑positive rate, with a detection limit of 0.2‑1.0 mg/L Hb. The pyramidon method delivers moderate sensitivity and moderate false‑positive rate, with a detection limit of 1‑5 mg/L Hb. Combined application of the two methods enables more reliable evaluation of occult bleeding. This kit is intended for research‑only use and shall not be used for clinical diagnosis or other purposes.
| Component No. | Component Name | Appearance | 2×100T | 2×300T | Storage |
| F1527137A | O-tolidine Solution | Liquid | 10ml | 30ml | 2-8℃.Store in the dark. |
| F1527137B | Pyramidon Solution | Liquid | 10ml | 30ml | 2-8℃.Store in the dark. |
| F1527137C | Oxidant | Liquid | 2×10ml | 2×30ml | 2-8℃.Store in the dark. |
Materials to Be Supplied by User
1. Fecal specimens, wooden spatula or toothpick, disinfected cotton pad or white porcelain plate, dropper or pipette
Operating Procedures (For Reference Only)
1. Collect fecal specimens and perform detection promptly. Prolonged storage will reduce reaction sensitivity; avoid water contamination.
2. Pick a small amount of feces (10‑50 mg, mung‑bean size) with wooden spatula or toothpick and smear it onto disinfected cotton pad or white porcelain plate.
3. o‑Tolidine method: add 2 drops (~0.1 mL) of o‑tolidine solution onto different positions of fecal smear. Pyramidon method: add 2 drops (~0.1 mL) of pyramidon solution onto different positions of fecal smear.
4. Add 2 drops (~0.1 mL) of oxidizing agent, start timing immediately and observe color change.
5. o‑Tolidine method: read result within 2 min. Blue color developing within 2 min indicates fecal hemoglobin (Hb) presence or positive FOBT result. Pyramidon method: read result within 2 min. Purple color developing within 2 min indicates fecal hemoglobin (Hb) presence or positive FOBT result.
6. Use reagents promptly after opening to prevent compromised experimental performance.
Result Interpretation
| Interpretation | Color Change (o‑Tolidine Method) | Color Change (Pyramidon Method) |
|---|---|---|
| (4+) | Immediate blue‑black‑brown color upon reagent addition | Immediate deep purple color upon reagent addition |
| (3+) | Immediate blue‑brown color, gradually turning black‑brown | Immediate purple color, gradually turning deep purple |
| (2+) | Initial pale‑green color, gradually developing distinct blue‑brown | Initial pale‑purple color, gradually developing distinct purple |
| (+) | Color shifts from pale‑green to green after 10 s | Color shifts from colorless to pale‑purple / purple after 10 s |
| Negative | No color development within 2 min | No color development within 2 min |
Precautions
1. During fecal specimen collection, transportation and reagent‑based detection, operators shall take adequate personal protective measures and avoid direct body contact to prevent contamination and self‑infection.
2. Trace gastrointestinal bleeding or irritation‑provoked bleeding may occur in normal humans and animals, which may lead to false‑positive results of this assay.
3. For suspected minor bleeding without obvious clinical symptoms, collect samples for at least 3 consecutive days and perform duplicate assays using material sampled from different sites of each specimen.
4. Subjects shall avoid animal blood, meat, liver, high‑chlorophyll foods, iron supplements, traditional Chinese medicines and other special medications for ≥3 days before sampling to prevent false‑positive reactions.
5. For experimental animals with colitis models, diet shall contain no fish meal, iron supplements or special medications to avoid false‑positive reactions.
6. o‑Tolidine Solution and Pyramidon Solution contain acetic acid with strong irritancy. Manipulate inside a fume hood. Solution color may change with storage time, which does not impair detection sensitivity.
7. Store oxidizing agent in sealed condition at 4 ℃. Its activity may decrease after 6 months. Commercially‑available medical hydrogen peroxide or urea peroxide can be used to prepare 1‑3 % working solution as alternative.
8. Gradient control experiments with blood or hemoglobin solution are recommended if necessary.
9. Do not store this kit together with oxidizing agents. Avoid direct sunlight exposure as well as excessively high or low‑temperature storage conditions.
10. Wear laboratory coat and disposable gloves for your safety and health.
Shelf‑Life
Valid for 12 months; transport under low‑temperature conditions, store at 4 ℃.
| 组分货号 | 组分名称 | 物理外观 | 2×100T | 2×300T | 储存 |
| F1527137A | 邻联甲苯胺溶液 | 液体 | 10ml | 30ml | 2-8℃.避光 |
| F1527137B | 匹拉米洞溶液 | 液体 | 10ml | 30ml | 2-8℃.避光 |
| F1527137C | 氧化剂 | 液体 | 2×10ml | 2×30ml | 2-8℃.避光 |
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