CN104372066A - 出血性大肠杆菌o157:h7选择性显色培养基及其检测试纸 - Google Patents

出血性大肠杆菌o157:h7选择性显色培养基及其检测试纸 Download PDF

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CN104372066A
CN104372066A CN201410681195.0A CN201410681195A CN104372066A CN 104372066 A CN104372066 A CN 104372066A CN 201410681195 A CN201410681195 A CN 201410681195A CN 104372066 A CN104372066 A CN 104372066A
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escherichia coli
enterohemorrhagic escherichia
selective
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金京勋
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SOOCHOW KH BIO SCIENCE&TECHNOLOGY Co Ltd (KHB)
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SOOCHOW KH BIO SCIENCE&TECHNOLOGY Co Ltd (KHB)
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Abstract

本发明提供一种出血性大肠杆菌O157:H7选择性显色培养基,其中每1000mL培养基含有如下重量的组分:结晶紫中性红胆盐琼脂15-25g、特异性酶显色底物5-25g、二甲基甲酰胺0.05-20g、酵母粉1-5g、山梨醇1-8g、头孢克肟15-25g、鼠李糖2-8g、亚碲酸钾2-7g、水杨苷1-5g、氯化钠5-10g、月桂基硫酸钠0.1-0.3g、万古霉素0.02-0.05g、余量为水。本发明还提供了一种含有所述出血性大肠杆菌O157:H7选择性显色培养基的检测试纸。本发明在CT-SMAC培养基基础上添加水杨苷、万古霉素,进一步提高选择性和特异性,检出率高、检出时间短。

Description

出血性大肠杆菌O157:H7选择性显色培养基及其检测试纸
技术领域
本发明涉及一种适用于出血性大肠杆菌O157:H7的选择性显色培养基,以及采用该选择性显色培养基的检测试纸。
背景技术
产志贺毒素大肠杆菌(shiga toxin-producing Escherichia coli,STEC)也称为产vero毒素大肠杆菌,是近来新出现的最重要的食源性病原菌。STEC主要引起胃肠炎和一系列并发症,如出血性腹泻(HC)和导致儿童肾衰竭的溶血性尿毒综合征(HUs)。肠出血性大肠杆菌(enterohaemorrhagic Escherichiacoli,EHEC)O157:H7是其中一种。反刍动物,特别是牛、山羊和绵羊已被认定是其重要宿主。它通过被污染的未煮熟的肉类、未消毒的牛奶和被粪便污染的蔬菜和水等传播,人-人之间的密切接触传播也有报道。大部分引起HC和HUS的爆发和散发病例与O157:H7有关。
O157可以迅速的分解乳糖,因此在麦康凯琼脂培养基上无法和其他大肠杆菌相区别。然而不像大约80%的其他大肠杆菌,大部分O157经过培养后,不分解山梨醇。因此利用这个特性建立了山梨醇-康凯琼脂(SMAC)培养方法,用山梨醇取代麦康凯琼脂中的乳糖,便可制备SMAC。已证明SMAC对分离O157是很有效的选择性培养基之一,目前被广泛用于实验室临床诊断。然而,近几年在世界各地发现了分解山梨醇的O157菌株,因此分离过程中应注意山梨醇发酵阳性菌株的挑取,以防漏检。
一些非O157血清型的大肠埃希菌也不发酵山梨醇;同时一些其他种属的非发酵山梨醇细菌也存在于人和动物的粪便中,特别是变形杆菌属和气单胞菌属细菌。基于这种情况,这就需要发展能抑制粪便中不发酵山梨醇的细菌生长的培养基。头孢克肟能抑制变形杆菌的生长而不抑制大肠杆菌,O157不发酵鼠李糖,而大部分不发酵山梨醇的大肠杆菌发酵鼠李糖。于是,人们就制备了头孢克肟-鼠李糖-山梨醇麦康凯培养基(CR-SMAC)。亚碲酸钾80年前就被用于病原菌的分离,包括棒状杆菌、白喉杆菌、金黄色葡萄球菌、霍乱弧菌和志贺氏菌。它对O157也具有选择性,于是发展了头孢克肟-亚碲酸钾-山梨醇麦康凯培养基(CT-SMAC)。然而,仍然有某些细菌能在CT-SMAC上形成与O157相似的菌落,使得在CT-SMAC平板上选择O157的困难增加。
发明内容
本发明提供一种适用于出血性大肠杆菌O157:H7的选择性显色培养基,以及采用该选择性显色培养基的检测试纸,以克服现有技术存在的上述缺陷。
本发明首先涉及一种出血性大肠杆菌O157:H7选择性显色培养基,其中每1000mL培养基含有如下重量的组分:
所述特异性酶显色底物是5-溴-4-氯-3-吲哚-α-D-葡萄糖苷。
本发明还涉及一种采用该选择性显色培养基制作的检测试纸,其至少含有一吸附有上述出血性大肠杆菌O157:H7选择性显色培养基的吸水滤纸。
本发明在CT-SMAC培养基基础上添加水杨苷、万古霉素,进一步提高选择性和特异性,检出率高、检出时间短。
具体实施方式
下面给出本发明较佳实施例,以详细说明本发明的技术方案。
实施例1
一种出血性大肠杆菌O157:H7选择性显色培养基,每1000mL培养基含有:结晶紫中性红胆盐琼脂15g、5-溴-4-氯-3-吲哚-α-D-葡萄糖苷5g、二甲基甲酰胺0.05g、酵母粉1g、氯化钠5g、月桂基硫酸钠0.3g、山梨醇1g、头孢克肟15g、鼠李糖2g、亚碲酸钾2g、水杨苷1g、万古霉素0.02g。
将上述成分溶于水至1000mL,加热煮沸,120℃灭菌15min,即可使用。
实施例2
一种出血性大肠杆菌O157:H7选择性显色培养基,每1000mL培养基含有:结晶紫中性红胆盐琼脂25g、5-溴-4-氯-3-吲哚-α-D-葡萄糖苷25g、二甲基甲酰胺20g、酵母粉5g、氯化钠10g、月桂基硫酸钠0.1g、月桂基硫酸钠0.3g、山梨醇8g、头孢克肟25g、鼠李糖8g、亚碲酸钾7g、水杨苷5g、万古霉素0.05g。
将上述成分溶于水至1000mL,加热煮沸,120℃灭菌15min,即可使用。
实施例3
一种出血性大肠杆菌O157:H7选择性显色培养基,每1000mL培养基含有:结晶紫中性红胆盐琼脂20g、5-溴-4-氯-3-吲哚-α-D-葡萄糖苷15g、二甲基甲酰胺5g、酵母粉3g、氯化钠8g、月桂基硫酸钠0.2g、山梨醇3g、头孢克肟20g、鼠李糖5g、亚碲酸钾3g、水杨苷2g、万古霉素0.03g。
将上述成分溶于水至1000mL,加热煮沸,120℃灭菌15min,即可使用。
实施例4
出血性大肠杆菌O157:H7选择性显色培养基试纸的制备:将选定的吸水滤纸浸入出血性大肠杆菌O157:H7选择性显色培养基中浸渍10-15分钟后取出烘干,冷却至室温后裁成一定规格试纸备用。
实施例5
在实施例4得到的试纸上下两面分别盖覆一层聚丙烯保护膜。
实施例6
将标准菌株分别制成100cfu/mL的标准菌悬液,吸取1mL菌液进行检测,37℃培养18小时。结果如表1,“+”表示阳性结果,“一”表示阴性结果。
表1 显色培养基特异性检验结果
由上表可以看出,除出血性大肠杆菌O157:H7以外的志贺氏菌属、弧菌、沙门氏菌属等主要食品致病菌在培养基上培养18h时进行观察,并没有菌落出现。特别是可选择性鉴别大肠埃希氏菌O26,具有十分重要的实用价值。可见本发明培养基对出血性大肠杆菌O157:H7初筛检测具有较好的选择性及特异性。

Claims (3)

1.一种出血性大肠杆菌O157:H7选择性显色培养基,其特征在于,其中每1000mL培养基含有如下重量的组分:
2.如权利要求1所述的出血性大肠杆菌O157:H7选择性显色培养基,其特征在于,所述特异性酶显色底物是5-溴-4-氯-3-吲哚-α-D-葡萄糖苷。
3.一种出血性大肠杆菌O157:H7选择性显色培养基检测试纸,其特征在于,其中至少含有一吸附有权利要求1或2所述出血性大肠杆菌O157:H7选择性显色培养基的吸水滤纸。
CN201410681195.0A 2014-11-24 2014-11-24 出血性大肠杆菌o157:h7选择性显色培养基及其检测试纸 Pending CN104372066A (zh)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108129530A (zh) * 2018-01-24 2018-06-08 广东优尼德生物科技有限公司 检测β-D-葡萄糖醛酸苷酶的底物及其制备方法和试剂盒
CN113293193A (zh) * 2021-06-01 2021-08-24 上海市食品药品检验研究院 一种检测大肠埃希菌o157的分离培养基及其制备方法

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
TOMOHIKO FUJISAWA等: "Modification of Sorbitol MacConkey Medium Containing Cefixime and Tellurite for Isolation of Escherichia coli O157:H7 from Radish Sprouts", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 *
刘斌主编: "《食品微生物检验》", 31 December 2013 *
卢勉飞等: "显色培养基快速检测大肠菌群和大肠杆菌效果的研究", 《中华预防医学杂志》 *
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108129530A (zh) * 2018-01-24 2018-06-08 广东优尼德生物科技有限公司 检测β-D-葡萄糖醛酸苷酶的底物及其制备方法和试剂盒
CN113293193A (zh) * 2021-06-01 2021-08-24 上海市食品药品检验研究院 一种检测大肠埃希菌o157的分离培养基及其制备方法

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Application publication date: 20150225