布氏杆菌 (Brucella) 是一种革兰氏阴性的不运动细菌，牛、羊、猪等动物最易感染，布氏杆菌染色又称科兹洛夫斯基染色或柯兹洛夫斯基染色。
阿拉丁布氏杆菌染色液采用沙黄为染色剂，临床标本直接涂片，背景干净，胞核胞质对比强烈，胞内吞噬体清晰易辨认，细菌染色特征典型，所有操作应置于二级生物安全柜中进行，一旦发现绿色细菌中有红色点状细菌，应高度怀疑为布氏杆菌，及时通知CDC，防止疫情扩散。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
产品组分及储存条件：
B1510429
| Component
| 2×50mL
| Storage
|
B1510429A
| 沙黄染色液
| 50mL
| RT
|
B1510429B
| 复染液
| 50mL
| RT |
自备材料：
接种环或挑取细菌的其他工具、酒精灯、载玻片、光学显微镜
操作步骤(仅供参考)：
1、涂片：取待检细菌，于载玻片中央涂成薄层，尽量薄；如果需做对照，可以将大肠杆菌与待检细菌混合涂片。
2、干燥：涂片后在室温下自然干燥，也可在酒精灯上略加温，使之迅速干燥。
3、固定：手持载玻片一端，标本面朝上，在酒精灯的火焰外侧快速来回移动3~5次，每次1s，温度不宜过高，防止菌体蛋白变性，放置待凉后染色。
4、染色：滴加沙黄染色液染色，在酒精灯的火焰上微微加热至出现蒸汽为止，通常需要30~60s。清水洗去染色液。
5、复染：滴加复染液染色2~3min。水洗，吸干，镜检。
染色结果：
注意事项：
1、涂片之前应事先在背面做好圆圈标记，以便判断后续试验的位置。
2、取细菌时，应注意自我防护，拔或塞试管塞时，应将试管口通过火焰略加烧灼，最后将接种环在火焰上烧灼灭菌。
3、加热固定涂片时，应注意玻片勿太靠近火焰，一般要求玻片温度不超过60°C，以玻片背面触及手背皮肤不觉过烫为宜。
4、待检细菌培养时间也会影响染色，阳性菌培养时间过长或已死亡或细菌溶解，都常呈阴性反应。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
6、试剂开封后请尽快使用，以防影响后续实验效果。
Brucella is a type of non-motile Gram-negative bacterium, with cattle, sheep, pigs and other animals being the most susceptible to infection. Brucella staining is also known as Kozlovsky staining.
Aladdin Brucella Stain Solution uses safranine as the stain. For direct smears of clinical specimens, it yields a clean background, strong contrast between cell nuclei and cytoplasm, and clear and distinguishable intracellular phagosomes, with typical bacterial staining characteristics. All operations should be performed in a Class II biosafety cabinet. If red dot-like bacteria are observed within green-stained bacteria, Brucella infection should be highly suspected. The CDC must be notified promptly to prevent the spread of the epidemic. This reagent is for research use only and is not suitable for clinical diagnosis or other purposes.
Product Components and Storage Conditions:
B1510429
| Component
| 2×50mL
| Storage
|
B1510429A
| Safranine Staining Solution
| 50mL
| RT
|
B1510429B
| Counterstain Solution
| 50mL
| RT |
Materials to Be Prepared:
Inoculation loop or other bacteria-picking tools, alcohol lamp, glass slides, optical microscope
Operating Procedures (For Reference Only):
1. Smear Preparation: Take the bacteria to be tested and spread them into a thin layer in the center of a glass slide; the thinner the better. If a control is required, mix Escherichia coli with the bacteria to be tested and prepare the smear together.
2. Drying: Air-dry the smear naturally at room temperature, or heat it gently over an alcohol lamp to speed up the drying process.
3. Fixation: Hold one end of the glass slide with the specimen side facing up, and move it quickly back and forth 3–5 times over the outer flame of an alcohol lamp, 1 second each time. Do not use excessive heat to avoid denaturation of bacterial proteins. Let the slide cool down before staining.
4. Staining: Add safranine staining solution to cover the smear, and heat it gently over an alcohol lamp until steam appears, which usually takes 30–60 seconds. Rinse the staining solution off with clean water.
5. Counterstaining: Add counterstain solution and stain for 2–3 minutes. Rinse with water, blot dry, and observe under a microscope.
Staining Results:
Brucella
| Red
|
Escherichia coli or other bacteria
| Green
|
Precautions:
1. Mark a circle on the back of the glass slide before preparing the smear to identify the position for subsequent tests.
2. Pay attention to personal protection when sampling bacteria. When removing or replacing the test tube stopper, pass the mouth of the test tube through the flame for brief sterilization. Finally, sterilize the inoculation loop by flaming it thoroughly.
3. When heating the slide for fixation, do not hold the slide too close to the flame. The temperature of the slide should generally not exceed 60°C, which can be judged by touching the back of the slide to the back of your hand—it should feel warm but not scalding.
4. The culture time of the bacteria to be tested can also affect the staining result. Positive bacteria that have been cultured for too long, are already dead, or have undergone lysis often show a negative staining reaction.
5. For your safety and health, wear a lab coat and disposable gloves during the operation.
6. Use the reagent as soon as possible after opening to avoid affecting the results of subsequent experiments.
ZJ26F0333470
危险品化学品经营许可证（带存储）