| 培养基名称 | 2%氯化钠碱性蛋白胨水，碱性蛋白胨水(ISO标准），ASPW培养基 |
|---|---|
| 英文名称: |
Alkaline Saline Peptone Water（ISO），ASPW |
| 培养基类型: | 选择性培养基 |
| 产品目录号: | M744 |
| 产品规格: |
250g，500g，5kg |
| 保存条件: | 密封，至于30度以下，干燥处保存。 |
| 产品性状: |
乳白色至黄色粉末。 浅黄色透明液体。 |
| 注意事项: | 避免摄入、呼入、皮肤接触。配制时在通风橱中进行，戴口罩、手套、护目镜。 |
| 相关产品: | 碱性蛋白胨水 |
产品描述：
ISO标准碱性蛋白胨水（Alkaline Saline Peptone Water）简称ASPW培养基,ISO标准弧菌增菌培养基，用于检测海鲜食品和临床样本中的弧菌。
本产品采用ISO标准配方。
2%碱性蛋白胨水工作原理
霍乱弧菌是引起霍乱疫情的成因。霍乱毒素（CT）是该菌的首要毒性因子。大多数从食品和环境中分离到的霍乱弧菌菌株不具有产毒能力，因此不是烈性的。多种生物化学特性和抗原性被作为该菌种的特征。摄入未经加热处理的海鲜引起的腹泻常常伴随拟态弧菌。分离弧菌应该做CT试验或检测CTX基因。经过富集后，筛选和验证该毒素可以用小鼠Y-1肾上腺细胞测定法和免疫测定法。
碱性蛋白胨水（ISO标准）最初作为非选择性气单孢菌属培养基。Cruickshank报导：如果提高pH值，该培养基可以有效地培养弧菌属。碱性蛋白胨水（ISO）利用了弧菌对高氯化钠浓度和碱性环境的耐受性。20%的氯化钠可以促进霍乱弧菌的生长。碱性条件能够抑制背景杂菌的生长。
ASPW培养基的用途：
ASPW培养基被APHA推荐用于检测食品和临床样本中的弧菌。
碱性蛋白胨水（ISO标准）也可直接用于弧菌的镜检。
ASPW培养基配方与配制方法
| 培养基成分： | 含量/L |
|---|---|
| 蛋白胨 | 20.0 g |
| 氯化钠 | 20.0 g |
| pH | 8.5 ± 0.2 |
配制方法：
1. 称取40g本品，加入1000ml去离子水溶解。
2. 分装到螺口盖试管中。
3. 121°C灭菌10min。
实验方法
弧菌的分离方法有多种，这些方法基本都涉及预增菌，然后接种到固体平板上，做形态学、生物化学和血清学鉴定。很多培养基被用作弧菌增菌培养基，但是这些培养基中仅碱性蛋白胨水（ISO标准）被广泛采用。
检测霍乱弧菌和副溶血弧菌
The following is an overview of ISO/TS 21872-1:20071 – Horizontal method for the detection of potentially enteropathogenic Vibrio spp. Part 1: Detection of Vibrio parahaemolyticus and Vibrio cholerae. Prepare samples in accordance with ISO 6887 (appropriate part)5, ISO 72186 and ISO 82617.
1. Add xg of sample to 9xml or 9xg of ASPW (ISO) at ambient temperature.
2. Incubate for 6 ± 1 hour, at 41.5 ± 1°C for fresh products or 37 ± 1°C for deep-frozen/salted products.
3. Remove 1ml of incubated medium and inoculate into a fresh 10ml of Alkaline Saline Peptone Water (ISO) and incubate for 18 ± 1 hour at 41.5°C.
4. From the cultures obtained in the ASPW (ISO), inoculate with a microbiological loop the surface of a TCBS Agar plate and second optional selective isolation medium in order to obtain single colonies.
5. Incubate the plates at 37°C for 24 ± 3 hours (or as appropriate for the second optional medium).
6. Examine the plates for the presence of typical colonies* of presumptive pathogenic Vibrio spp. Select 5 typical colonies for subculture onto Saline Nutrient Agar followed by biochemical confirmation.
其它弧菌的检测
The following is an overview of ISO/TS 21872-2:20072 – Horizontal method for the detection of potentially enteropathogenic Vibrio spp. Part 2: Detection of species other than Vibrio parahaemolyticusand Vibrio cholerae. Prepare samples in accordance with ISO 6887 (appropriate part)5, ISO 72186 and ISO 82617.
1. Add xg of sample to 9xml or 9xg of ASPW (ISO) at ambient temperature.
2. Incubate for 6 ± 1 hour, at 41.5- ± 1°C for fresh products or 37 ± 1°C for deep-frozen/salted products.
3. Remove 1 ml of incubated medium and inoculate into a fresh 10ml of Alkaline Saline Peptone Water (ISO) and incubate for 18 ± 1 hour at 37 ± 1°C.
4. From the cultures obtained in the ASPW (ISO), inoculate with a microbiological loop the surface of a TCBS Agar plate and second optional selective isolation medium in order to obtain single colonies.
5. Incubate the plates at 37°C for 24 ± 3 hours (or as appropriate for the second optional medium).
6. Examine the plates for the presence of typical colonies† of presumptive pathogenic Vibrio spp. Select 5 typical colonies for subculture onto Saline Nutrient Agar followed by biochemical confirmation.
结果与分析
标准菌株 生长结果
Vibrio parahaemolyticus NCTC10885 浓密生长
Vibrio furnissii NCTC11218 浓密生长
Vibrio vulnificus ATCC29307 浓密生长
Vibrio cholerae non-O1/non-O139 ATCC14733 浓密生长
