CN108103139A - 一种沙门菌的快速检测方法 - Google Patents
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Abstract
一种沙门菌的快速检测方法,包括以下步骤:S1、样品处理:称取固体样品2‑5g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;S2、配制培养基;S3、培养基冷却至室温后,对培养基进行紫外灭菌15‑20min;S4、细菌培养:将待测样品置入培养基中培养4‑6h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液。本发明具有操作步骤简单、方便的优点,周期较短,能够快速对沙门菌进行检测,节约了检测时间,便于实际检测使用。
Description
技术领域
本发明涉及沙门菌检测方法技术领域,尤其涉及一种沙门菌的快速检测方法。
背景技术
沙门氏菌是一种严重威胁人类及动物生命健康的人畜共患病原菌,可引起人和动物急性胃肠炎、伤寒、败血症以及肠外灶性感染等,已被FAO列为A类病原微生物。历年来,沙门氏菌引起的食物中毒常列于细菌性食物中毒的榜首。我国疾病预防控制中心已将沙门氏菌列为食品中要求控制的细菌指标之一。准确及时的检测手段是预防和控制病原微生物传播的关键。目前沙门氏菌的检测方法通常都过于繁琐且周期交长,通常需要很多天才能完成检测工作,不便于对沙门氏菌进行快速检测。
发明内容
本发明的目的是为了解决现有技术中存在的不便于对沙门氏菌进行快速检测的缺点,而提出的一种沙门菌的快速检测方法。
为了实现上述目的,本发明采用了如下技术方案:
设计一种沙门菌的快速检测方法,包括以下步骤:
S1、样品处理:称取固体样品2-5g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌15-20min;
S4、细菌培养:将待测样品置入培养基中培养4-6h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.2±0.2,再加入孔雀绿36mg,进行分装、灭菌,备用;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养6-10h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加3-4滴至胶体金检测卡的样品孔,反应5-10min,判读结果。
优选的,所述步骤S5中的灭菌温度不超过115℃。
优选的,所述步骤S2中的培养基设置为BPW培养基。
本发明提出的一种沙门菌的快速检测方法,有益效果在于:本发明具有操作步骤简单、方便的优点,周期较短,能够快速对沙门菌进行检测,节约了检测时间,便于实际检测使用。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。
一种沙门菌的快速检测方法,包括以下步骤:
S1、样品处理:称取固体样品2g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基,培养基设置为BPW培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌15min;
S4、细菌培养:将待测样品置入培养基中培养4h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.0,再加入孔雀绿36mg,进行分装、灭菌,备用,灭菌温度不超过115℃;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养6h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加3滴至胶体金检测卡的样品孔,反应5min,判读结果。
实施例2
一种沙门菌的快速检测方法,包括以下步骤:
S1、样品处理:称取固体样品4g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基,培养基设置为BPW培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌17min;
S4、细菌培养:将待测样品置入培养基中培养5h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.2,再加入孔雀绿36mg,进行分装、灭菌,备用,灭菌温度不超过115℃;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养7h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加3滴至胶体金检测卡的样品孔,反应6min,判读结果。
实施例3
一种沙门菌的快速检测方法,包括以下步骤:
S1、样品处理:称取固体样品3g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基,培养基设置为BPW培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌20min;
S4、细菌培养:将待测样品置入培养基中培养5h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.2,再加入孔雀绿36mg,进行分装、灭菌,备用,灭菌温度不超过115℃;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养7h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加4滴至胶体金检测卡的样品孔,反应8min,判读结果。
实施例4
一种沙门菌的快速检测方法,包括以下步骤:
S1、样品处理:称取固体样品5g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基,培养基设置为BPW培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌20min;
S4、细菌培养:将待测样品置入培养基中培养6h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.4,再加入孔雀绿36mg,进行分装、灭菌,备用,灭菌温度不超过115℃;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养10h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加4滴至胶体金检测卡的样品孔,反应10min,判读结果。
以上所述,仅为本发明较佳的具体实施方式,但本发明的保护范围并不局限于此,任何熟悉本技术领域的技术人员在本发明揭露的技术范围内,根据本发明的技术方案及其发明构思加以等同替换或改变,都应涵盖在本发明的保护范围之内。
Claims (3)
1.一种沙门菌的快速检测方法,其特征在于,包括以下步骤:
S1、样品处理:称取固体样品2-5g并粉碎,在粉碎后的固体样品中加入无菌生理盐水进行溶解;
S2、配制培养基;
S3、培养基冷却至室温后,对培养基进行紫外灭菌15-20min;
S4、细菌培养:将待测样品置入培养基中培养4-6h,将培养后的产物进行细菌鉴定,取鉴定后的菌落加入无菌生理盐水配制成菌悬液;
S5、增菌:配制增菌培养基:取大豆胨4.5g、六水合氯化镁29.0g、氯化钠8.0g、磷酸氢二钾0.4g、蒸馏水1L、磷酸二氢钾0.6g进行混合,微温溶解,调节pH值,并使灭菌后的增菌培养基在25℃的pH值为5.2±0.2,再加入孔雀绿36mg,进行分装、灭菌,备用;
S6、b1、取25g样品菌悬液到均质袋中,并向其中加入200mL的增菌培养基,放入均质器中均质3min;
b2、将均质后的溶液在42℃的环境下静止培养6-10h;
S7、将培养物混合均匀,并取1mL到试管中沸水加热10min,冷却至室温,用滴管滴加3-4滴至胶体金检测卡的样品孔,反应5-10min,判读结果。
2.根据权利要求1所述的一种沙门菌的快速检测方法,其特征在于,所述步骤S5中的灭菌温度不超过115℃。
3.根据权利要求1所述的一种沙门菌的快速检测方法,其特征在于,所述步骤S2中的培养基设置为BPW培养基。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
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US20040038197A1 (en) * | 2002-08-22 | 2004-02-26 | Usda Ars Ott | Extraction and concentration method |
CN105062921A (zh) * | 2015-08-12 | 2015-11-18 | 华南农业大学 | 一种高效抑制禽致病性沙门氏菌的唾液乳杆菌及其应用 |
CN106896225A (zh) * | 2017-04-28 | 2017-06-27 | 河南艾能生物科技有限公司 | 一种沙门氏菌的快速检测方法 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20040038197A1 (en) * | 2002-08-22 | 2004-02-26 | Usda Ars Ott | Extraction and concentration method |
CN105062921A (zh) * | 2015-08-12 | 2015-11-18 | 华南农业大学 | 一种高效抑制禽致病性沙门氏菌的唾液乳杆菌及其应用 |
CN106896225A (zh) * | 2017-04-28 | 2017-06-27 | 河南艾能生物科技有限公司 | 一种沙门氏菌的快速检测方法 |
Non-Patent Citations (3)
Title |
---|
刘富来等: ""鸭肠炎沙门氏菌分离鉴定与药敏试验"", 《动物医学进展》 * |
焦新安主编: "《食品检验检疫学》", 30 January 2007, 中国农业出版社 * |
赫士海主编: "《现代细菌学培养基和生化试验手册》", 28 February 1992, 中国科学技术出版社 * |
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