Oxoid培养基 CM0910B MOD SEMI-SOLID RAP VASS(MSRV)M使用说明

Oxoid培养基 CM0910B MOD SEMI-SOLID RAP VASS(MSRV)M使用说明

Dehydrated Culture Media

MODIFIED SEMI-SOLID RAPPAPORT VASSILIADIS (MSRV) MEDIUM BASE

改良型半固态拉帕波特 VASSILIADIS (MSRV) 中型底座

Code: CM0910

A semi-solid medium for the detection of motile Salmonella species from food and environmental samples.

一种半固体培养基,用于检测食品和环境样品中的运动沙门氏菌。

Typical Formula*

gm/litre

Tryptose

4.59

Casein hydrolysate

4.59

Sodium chloride

7.34

Potassium dihydrogen phosphate

1.47

Magnesium chloride (anhydrous)

10.93

Malachite green oxalate

0.037

Agar

2.7

pH 5.4 ± 0.2 @ 25°C
* Adjusted as required to meet performance standards

MSRV SELECTIVE SUPPLEMENT  MSRV 选择性补充

Code: SR0161

货号:SR0161

Vial contents (each vial is sufficient for 500ml of medium)
per vial
per litre
Novobiocin

10.0mg

20.0mg

Directions
Suspend 15.8g of MSRV Medium Base in 500ml of distilled water. Bring to the boil with frequent agitation.
DO NOT AUTOCLAVE. Cool to 50°C and aseptically add the contents of 1 vial of MSRV Selective Supplement SR0161E reconstituted with 2ml of sterile distilled water. Mix well and pour into sterile Petri dishes. Air dry at room temperature for at least one hour. (Plates may be air-dried overnight prior to storage at 2-8°C.)

Description
Modified Semi-solid Rappaport Vassiliadis (MSRV) Medium is based on the formulation described by De Smedt et al. which has been shown to detect more Salmonella-positive samples than the traditional enrichment procedures1,2. Further collaborative studies have confirmed these findings3,4.

Motility enrichment on MSRV Medium has been designed as a simple, sensitive method for the isolation of salmonellae from food and environmental samples. The efficiency of the medium is based on the ability of salmonellae to migrate through the selective medium ahead of competing motile organisms, thus producing opaque halos of growth.

Further tests can be carried out directly from the migrated culture with the inoculum being taken from the edge of the growth. Oxoid Salmonella Latex Test FT0203 or DR1108 may be used for serological confirmation of Salmonella species.

The medium is not suitable for the detection of non-motile strains of Salmonella (incidence5. (Figures obtained from records of the Department of Enteric Pathogens, Central Public Health Laboratory, Colindale, London. Dr. B. Rowe, Personal Communication, 1988.)

Technique
1. Inoculate up to three drops (ca. 0.1ml) of the pre-enrichment culture (after incubation for 16-20 hours) in separate, evenly spaced spots on the surface of the MSRV Medium plates.
2. Incubate the plates in an upright position at 42°C for up to 24 hours. (Care should be taken not to exceed 24 hours.)
3. Examine the plates for motile bacteria which will be shown by a halo of growth originating from the inoculation spot.
4. Subcultures can be taken from the outside edge of the halo to confirm purity and for further biochemical and serological tests.

De Smedt6 reported that if MSRV medium is contained in test tubes and incubation is carried out under anaerobic conditions, visible migration zones are produced in 6 hours enabling Salmonella in foods to be detected in 24 hours.

Storage conditions and Shelf life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the selective supplement in the dark at 2-8°C and use before the expiry date on the label.
The prepared medium may be stored for up to 2 weeks at 2-8°C away from the light.

Appearance
Dehydrated medium: Green coloured, free-flowing powder
Prepared medium: Blue coloured semi-solid gel

方向
将 15.8 克 MSRV 培养基基质悬浮在 500 毫升蒸馏水中。经常搅拌煮沸。
不要高压灭菌。冷却至 50°C 并在无菌条件下加入 1 瓶 MSRV 选择性补充剂 SR0161E 中的内容物,该补充剂用 2ml 无菌蒸馏水复溶。充分混合并倒入无菌培养皿中。在室温下风干至少一小时。 (板可以在 2-8°C 储存前风干过夜。)

描述
改良的半固体 Rappaport Vassiliadis (MSRV) 培养基基于 De Smedt 等人描述的配方。与传统的富集程序相比,它已被证明可以检测到更多的沙门氏菌阳性样本 1,2。进一步的合作研究证实了这些发现3,4。

MSRV 培养基上的运动富集被设计为一种简单、灵敏的方法,用于从食品和环境样品中分离沙门氏菌。培养基的效率基于沙门氏菌在竞争性运动生物之前通过选择性培养基迁移的能力,从而产生不透明的生长晕圈。

进一步的测试可以直接从迁移的培养物中进行,接种物取自生长的边缘。氧化型沙门氏菌乳胶测试 FT0203 或 DR1108 可用于沙门氏菌属的血清学确认。

该培养基不适用于检测非运动型沙门氏菌菌株(发病率5。(数据来自伦敦科林代尔中央公共卫生实验室肠道病原体部的记录。B. Rowe 博士,个人通讯,1988 年。 )

技术
1. 在 MSRV 培养基板表面上的独立、均匀分布的点中接种最多三滴(约 0.1 毫升)预富集培养物(孵育 16-20 小时后)。
2. 将板在 42°C 直立位置孵育长达 24 小时。 (注意不要超过 24 小时。)
3. 检查平板上的活动细菌,这些细菌会以源自接种点的生长晕圈显示。
4. 可以从光环的外边缘进行亚培养,以确认纯度并进行进一步的生化和血清学测试。

De Smedt6 报告说,如果试管中包含 MSRV 培养基并在厌氧条件下进行孵育,则 6 小时内会产生可见的迁移区,从而可以在 24 小时内检测到食品中的沙门氏菌。

储存条件和保质期
将脱水培养基储存在 10-30°C 并在标签上的有效期之前使用。
将选择性补充剂储存在 2-8°C 的黑暗中,并在标签上的有效期之前使用。
制备好的培养基可以在 2-8°C 下避光保存长达 2 周。

外貌
脱水介质:绿色、自由流动的粉末
制备培养基:蓝色半固体凝胶

Quality control

Positive controls: Expected results
Salmonella poona NCTC 4840 Straw colonies at site of inoculation surrounded by halo of growth
Salmonella enteritidis ATCC® 13076 * Straw colonies at site of inoculation surrounded by halo of growth
Negative control:
Citrobacter freundii ATCC® 8090 * Restricted or no growth
* This organism is available as a Culti-Loop®

Precautions
The basal medium is very hygroscopic. When handling the powder a face mask and gloves must be worn, refer to the material safety data sheet.

References
1. De Smedt J. M., Bolderdijk R., Rappold H. and Lautenschlaeger D. (1986) J. Food. Prot. 49. 510-514.
2. De Smedt J. M., Bolderdijk R. (1987) J. Food. Prot. 50. 658-661.
3. De Zutter L. et al. (1991) Int. J. Food Micro. 13. 11-20.
4. De Smedt J. M. et al. (1991) Int. J. Food Micro. 13. 301-308.
5. Holbrook R., Anderson J. M., Baird-Parker A. C., Dodds L. M., Sawhney D., Struchbury S. H. and Swaine D. (1989) Lett. Appl. Microbiol. 8. 139- 142.
6. De Smedt, J. M. (1998) Abstract 1.5. Int. Symposium – Food borne Pathogens: Detection and Typing. The Hague, The Netherlands 20th-21st April 1998.

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