CN105062934A - 一种阪崎肠杆菌显色培养基 - Google Patents

一种阪崎肠杆菌显色培养基 Download PDF

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CN105062934A
CN105062934A CN201510580890.2A CN201510580890A CN105062934A CN 105062934 A CN105062934 A CN 105062934A CN 201510580890 A CN201510580890 A CN 201510580890A CN 105062934 A CN105062934 A CN 105062934A
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culture medium
sodium
enterobacter
enterobacter sakazakii
peptone
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刘士旺
楼坚
斯丹丹
施丽洁
周亚琴
尹学辉
方文姬
林学海
吴元锋
黄�俊
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Zhejiang Lover Health Science and Technology Development Co Ltd
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Abstract

一种阪崎肠杆菌显色培养基，属于生物技术领域。该培养基由以下组分构成：蛋白胨10-20g/L、酵母粉5-7g/L、氯化钠5-8g/L、柠檬酸铁铵0.5-1.5g/L、硫代硫酸钠0.5-1.5g/L、脱氧胆酸钠0.5-1.52g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.01-0.25g/L、琼脂13-20g/L，pH？6.8±0.2。本发明培养基组方简单，操作方便；实验结果便于观察，分析；提高检测效率和准确率；与传统培养基相比，其成本更低，利于推广应用。

Description

一种阪崎肠杆菌显色培养基
技术领域
本发明属于生物技术领域，具体涉及一种阪崎肠杆菌显色培养基。
背景技术
阪崎肠杆菌是肠杆菌科的一种有周生鞭毛、能运动、无芽孢、兼性厌氧的革兰阴性杆菌，以前被称为黄色阴沟肠杆菌，1980年更名为阪崎肠杆菌。该菌是人和动物肠道中的一种正常寄生菌，但是在一定条件下会致病。1961年，两位英国医生Urmenyi和Franklin首次报道了两例由阪崎肠杆菌引起的新生儿脑膜炎，随后丹麦、美国、荷兰、希腊、冰岛、比利时等国家相继报道了新生儿阪崎肠杆菌感染事件。新生儿感染该菌可能导致脑膜炎、坏死性小肠结肠炎和菌血症等疾病，并可引起严重的神经系统后遗症甚至死亡，感染致死率高达20%～50%，即使治愈，也容易造成神经系统性后遗症。该菌偶尔还可引起成人局部感染、菌血症以及骨髓炎等。阪崎肠杆菌已经被国际食品微生物标准委员会（ICMSF）称为是严重危害特定人群、威胁生命或导致慢性实质性后遗症的细菌。目前有关阪崎肠杆菌的污染来源尚不清楚。Leclercq等2002年从某些食品样本（例如奶酪、香料、碎牛肉等）中，Hamilton等2003年从厩螫蝇中均分离出了该菌，但从已经报道的诸多新生儿感染病例报告中基本可以确定婴儿配方奶粉是新生儿感染的主要途径。
显色培养基（Chromogenic/FluorogenicCultureMedia）是一类利用微生物自身代谢产生的酶与相应显色底物反应显色的原理来检测微生物的新型培养基。这些相应的显色底物是由产色基因和微生物部分可代谢物质组成，在特异性酶作用下，游离出产色基因显示一定颜色，直接观察菌落颜色即可对菌种作出鉴定。它是一种新型分离培养基，利用显色培养基进行微生物的筛选分离，其反应的灵敏度和特异性大大优于传统培养基。
传统的阪崎肠杆菌显色培养基中的显色剂为4-硝基苯酚-α-D-吡喃葡萄糖苷。4-硝基苯酚-α-D-吡喃葡萄糖苷产生的4-硝基苯酚在琼脂中很容易扩散影响实验结果观察，因而存在一定的局限性。此外，在TSA上加入4-甲基伞形花内酮-α-D葡萄糖苷酸（α-MUG）可鉴别阪崎肠杆菌疑似菌落，但其只能在紫外线照射条件下才能区分，且紫外线对人体有害。
发明内容
针对现有技术存在的问题，本发明的目的在于设计提供一种阪崎肠杆菌显色培养基的技术方案。
所述的一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨10-20g/L、酵母粉5-7g/L、氯化钠5-8g/L、柠檬酸铁铵0.5-1.5g/L、硫代硫酸钠0.5-1.5g/L、脱氧胆酸钠0.5-1.52g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.01-0.25g/L、琼脂13-20g/L，pH6.8±0.2。
所述的一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨12-18g/L、酵母粉5-7g/L、氯化钠6-7g/L、柠檬酸铁铵0.8-1.2g/L、硫代硫酸钠0.8-1.2g/L、脱氧胆酸钠0.8-1.3g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.015-0.2g/L、琼脂14-18g/L。
所述的一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨20g/L、酵母粉6g/L、氯化钠5g/L、柠檬酸铁铵1g/L、硫代硫酸钠1g/L、脱氧胆酸钠1g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.1g/L、琼脂15g/L。
本发明的积极效果：本发明培养基组方简单，操作方便；实验结果便于观察，分析；提高检测效率和准确率；与传统培养基相比，其成本更低，利于推广应用。
具体实施方式
以下结合实施例来进一步说明本发明。
实施例1
1、制备阪崎肠杆菌显色培养基，该培养基配方为：蛋白胨20.0g、酵母粉6.0g、氯化钠5.0g、柠檬酸铁铵1.0g、硫代硫酸钠1.0g、脱氧胆酸钠1.0g、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.1g、琼脂15g、pH6.8±0.2、蒸馏水1000ml。
称取6.0g该显色培养基，加入150ml蒸馏水，加热煮沸完全溶解，121℃高压灭菌15min，冷至50℃左右时，倾入无菌平皿，备用。
2、接种：将活化后的阪崎肠杆菌、枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、变形杆菌、鼠伤寒沙门氏菌、蜡样芽孢杆菌分别涂布于上述显色培养基，37℃培养24h。
3、结果：
实施例2
1、制备阪崎肠杆菌显色培养基，该培养基配方为：蛋白胨10g、酵母粉5g、氯化钠5g、柠檬酸铁铵0.5g、硫代硫酸钠0.5g、脱氧胆酸钠0.5g、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.01g、琼脂13g、pH6.8±0.2、蒸馏水1000ml。
称取6.0g该显色培养基，加入150ml蒸馏水，加热煮沸完全溶解，121℃高压灭菌15min，冷至50℃左右时，倾入无菌平皿，备用。
2、接种：将活化后的阪崎肠杆菌、枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、变形杆菌、鼠伤寒沙门氏菌、蜡样芽孢杆菌分别涂布于上述显色培养基，37℃培养24h。
3、结果：
实施例3
1、制备阪崎肠杆菌显色培养基，该培养基配方为：蛋白胨20g、酵母粉7g、氯化钠8g、柠檬酸铁铵1.5g、硫代硫酸钠1.5g、脱氧胆酸钠1.52g、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.25g、琼脂20g、pH6.8±0.2、蒸馏水1000ml。
称取6.0g该显色培养基，加入150ml蒸馏水，加热煮沸完全溶解，121℃高压灭菌15min，冷至50℃左右时，倾入无菌平皿，备用。
2、接种：将活化后的阪崎肠杆菌、枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、变形杆菌、鼠伤寒沙门氏菌、蜡样芽孢杆菌分别涂布于上述显色培养基，37℃培养24h。
3、结果：
实施例4
1、所述的阪崎肠杆菌显色培养基，其培养基配方为：蛋白胨20.0g、酵母粉6.0g、氯化钠5.0g、柠檬酸铁铵1.0g、硫代硫酸钠1.0g、脱氧胆酸钠1.0g、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.1g、琼脂15g、pH6.8±0.2、蒸馏水1000ml。
称取6.0g该显色培养基，加入150ml蒸馏水，加热煮沸完全溶解，121℃高压灭菌15min，冷至50℃左右时，倾入无菌平皿，备用。
2、接种：用接种环挑取阪崎肠杆菌、枯草芽孢杆菌、大肠杆菌、金黄色葡萄球菌、变形杆菌、鼠伤寒沙门氏菌、蜡样芽孢杆菌的单菌落于缓冲蛋白胨水（BPW）培养基混合活化。将混合培养液稀释10-4倍，再取100ul涂布于上述显色培养基，37℃培养24h。
3、结果：培养基中长出紫红色和灰黑色单菌落，紫红色单菌落鉴定为阪崎肠杆菌菌落。

Claims (3)

1.一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨10-20g/L、酵母粉5-7g/L、氯化钠5-8g/L、柠檬酸铁铵0.5-1.5g/L、硫代硫酸钠0.5-1.5g/L、脱氧胆酸钠0.5-1.52g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.01-0.25g/L、琼脂13-20g/L，pH6.8±0.2。
2.如权利要求1所述的一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨12-18g/L、酵母粉5-7g/L、氯化钠6-7g/L、柠檬酸铁铵0.8-1.2g/L、硫代硫酸钠0.8-1.2g/L、脱氧胆酸钠0.8-1.3g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.015-0.2g/L、琼脂14-18g/L。
3.如权利要求1所述的一种阪崎肠杆菌显色培养基，其特征在于由以下组分构成：蛋白胨20g/L、酵母粉6g/L、氯化钠5g/L、柠檬酸铁铵1g/L、硫代硫酸钠1g/L、脱氧胆酸钠1g/L、5-溴-6-氯-3-吲哚-α-D-吡喃葡萄糖0.1g/L、琼脂15g/L。
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105861623A (zh) * 2016-04-25 2016-08-17 无锡市赛微生物技术有限公司 一种用于检测阪崎肠杆菌的显色培养基
CN113416765A (zh) * 2021-08-12 2021-09-21 合肥工业大学 一种婴幼儿配方奶粉中克罗诺杆菌的检测方法

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105861623A (zh) * 2016-04-25 2016-08-17 无锡市赛微生物技术有限公司 一种用于检测阪崎肠杆菌的显色培养基
CN113416765A (zh) * 2021-08-12 2021-09-21 合肥工业大学 一种婴幼儿配方奶粉中克罗诺杆菌的检测方法
CN113416765B (zh) * 2021-08-12 2022-08-16 合肥工业大学 一种婴幼儿配方奶粉中克罗诺杆菌的检测方法

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Application publication date: 20151118