Oxoid CM0995B 洋葱博克霍尔德菌培养基 BURKHOLDERIA CEPACIA AGAR BASE

Oxoid CM0995B 洋葱博克霍尔德菌培养基 BURKHOLDERIA CEPACIA AGAR BASE

BURKHOLDERIA CEPACIA AGAR BASE

CODE: CM0995

A medium for the selective isolation of Burkholderia cepacia from the respiratory secretions of patients with cystic fibrosis and for routine testing of non-sterile inorganic salt solutions containing preservative.

一种培养基,用于从囊性纤维化患者的呼吸道分泌物中选择性分离洋葱伯克霍尔德菌,并用于常规检测含有防腐剂的非无菌无机盐溶液。

Typical Formula*

gm/litre

Peptone

5.0

Yeast Extract

4.0

Sodium pyruvate

7.0

Potassium dihydrogen phosphate

4.4

Disodium hydrogen phosphate

1.4

Bile salts

1.5

Ammonium sulphate

1.0

Magnesium sulphate

0.2

Ammonium ferrous sulphate

0.01

Phenol red

0.02

Crystal violet

0.001

Agar

12.0

pH 6.2 ± 0.2 @ 25°C
* Adjusted as required to meet performance standards

BURKHOLDERIA CEPACIA SELECTIVE SUPPLEMENT

CODE: SR0189

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

75,000IU

150,000IU

Gentamicin

2.5mg

5.0mg

Ticarcillin

50.0mg

100.0mg

Directions
Suspend 18.25g of Burkholderia cepacia Agar Base in 500ml of distilled water, mix well and sterilise by autoclaving at 121°C for 15 minutes. Cool to 50°C and aseptically add the contents of one vial of Burkholderia cepacia Selective Supplement SR0189, reconstituted as directed. Mix well and distribute into sterile Petri dishes.

Description
Burkholderia cepacia (formerly known as Pseudomonas cepacia) is a motile aerobic oxidase positive Gram-negative bacillus commonly found in liquid reservoirs and moist environments. The cells are 0.5 to 1.0mm wide and 5mm in length. It is an important opportunistic pathogen and causes pulmonary infection among individuals with cystic fibrosis (CF). Isolates from CF patients often display multidrug resistance and as many as 20% of colonised individuals will succumb to Burkholderia cepacia syndrome, a necrotizing pneumonia associated with fever that culminates into a rapid and fatal clinical deterioration1. Oxoid’s Burkholderia cepacia Agar is based on PC Medium originally devised by Gilligan et al. where it was shown to be superior for growth of Burkholderia cepacia after 48 hours when compared to MacConkey2.

Originally isolated from onions, Burkholderia cepacia can survive for long periods and multiply in hostile environments such as antiseptic and disinfectant solutions, distilled water, whirlpool baths, nebulizers and commercially packaged urinary catheter kits3. An outbreak in Arizona in 1998 due to contaminated alcohol-free mouthwash, was investigated by the Food and Drug Administration (FDA), who suggested an association with the deionisation procedure of the water used to prepare the product4. The organism may be present in low numbers in many non-sterile products used in hospitals. It has been isolated from various water sources and can grow in distilled water with a nitrogen source due to its ability to fix carbon dioxide from air5. Suction catheters rinsed in acetic acid solution have reduced incidence of transmission of Burkholderia cepacia and other pseudomonads.

The slower growing Burkholderia cepacia can be missed on conventional media such as blood or MacConkey Agar due to overgrowth caused by other faster growing organisms found in the respiratory tract of CF patients such as mucoid Klebsiella species, Pseudomonas aeruginosa and Staphylococcus species. This may lead to the infection being missed or wrongly diagnosed.

Technique
Take a routine respiratory sample from the patient e.g. sputa, deep pharyngeal swabs or bronchial washings. Dilute the sample, if necessary, in Ringer’s solution to give a 1:2 dilution. Streak onto Burkholderia cepacia Medium and incubate at 37°C for 48 to 72 hours.
Examine after 48 hours for sage green colonies and the medium turning from straw-green to bright pink. All colonies should be further identified and confirmed. Re-incubate for a further 24 hours if necessary.
Typical colonies of Burkholderia cepacia are circular, and entire. Colour formation is based on natural pigment expression and colonies vary from grey to sage green, with the medium changing from orange to bright pink.

Storage conditions and Shelf Life
Store the dehydrated medium at 10-30°C and use before the expiry date on the label.
Store the prepared plates of medium at 2-8°C.

Appearance
Dehydrated medium: Straw pink, free-flowing powder
Prepared medium: Orange coloured gel

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。
将准备好的培养基板储存在 2-8°C。

外貌
脱水介质:稻草粉色,自由流动的粉末
制备培养基:橙色凝胶



Quality control

Positive control: Expected results
Burkholderia cepacia ATCC® 25608* Good growth, grey colonies with bright pink medium. Low numbers of colonies may not produce a colour change of the medium.
Burkholderia cepacia ATCC® 25416* Good growth, sage green colonies with bright pink medium. Low numbers of colonies may not produce a colour change of the medium.
Negative control:
Pseudomonas aeruginosa ATCC® 27853* Inhibited
* This organism is available as a Culti-Loop®

References
1
.Whitby, P. W. (1998) Journal of Clinical Microbiology 36: 1642-1645
2. Gilligan, P. H. et al. (1985) Journal of Clinical Microbiology 22: No.1 5-8.
3. Geftic, S. G., Heymann, H. and Adair, F. W. (1979) Applied and Environmental Microbiology 37: 505-510
4. Matrician, L. (1998) Virtual Hospital: Morbidity and Mortality Weekly Report Volume 47: No. 43
5. Koneman, E. W. et al. (1997) Color Atlas and Textbook of Diagnostic Microbiology Fifth Ed.: 269-272

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