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用于从粪便中分离弯曲杆菌属的改良选择性培养基:与普雷斯顿培养基(一种无血培养基)及过滤系统的比较

Modified selective medium for isolation of Campylobacter spp. from feces: comparison with Preston medium, a blood-free medium, and a filtration system.

作者信息

Goossens H, De Boeck M, Coignau H, Vlaes L, Van den Borre C, Butzler J P

出版信息

J Clin Microbiol. 1986 Nov;24(5):840-3. doi: 10.1128/jcm.24.5.840-843.1986.

Abstract

Our previously described (H. Goossens, M. De Boeck, and J. P. Butzler, Eur. J. Clin. Microbiol. 2:389-393, 1983) selective medium, consisting of cefoperazone (15 mg/liter), rifampin (10 mg/liter), colistin (10,000 IU/liter), and amphotericin B (2 mg/liter) (medium M1), for the isolation of Campylobacter jejuni and Campylobacter coli from stool specimens was modified as follows: cefoperazone (30 mg/liter), rifampin (10 mg/liter), and amphotericin B (2 mg/liter) (medium M2). A comparative study of the isolation of Campylobacter spp. from stool specimens was carried out with medium M1; medium M2; a selective blood-free medium consisting (per liter) of charcoal (4 g), ferrous sulfate (0.25 g), sodium pyruvate (0.25 g), casein hydrolysate (3 g), sodium deoxycholate (1 g), nutrient broth no. 2 (25 g), agar (12 g), and cefoperazone (32 mg) (medium M3); and Preston medium containing (per liter) trimethoprim (10 mg), rifampin (10 mg), polymyxin B (5,000 IU), and cycloheximide (100 mg) (medium M4). We also included a filtration system in which membrane filters were applied directly to the surface of the nonselective blood-free medium distributed in small petri dishes. A total of 5,276 stool specimens were tested: 2,788 stool specimens were tested on M1 and M3 in study 1; 2,488 stool specimens were inoculated on the four selective media in study 2, and the last 986 specimens of the 2,488 were tested in parallel with the filtration system. In study 2, 128 Campylobacter strains were isolated from 126 different patients; 85.0, 88.3, 82.5, and 66.6% of these strains were isolated on M1, M2, M3, and M4, respectively. No contaminating fecal flora was found on 65.4, 70.7, 62.4, and 40.3% of the M1, M2, M3, and M4 plates, respectively. Furthermore, C. coli was found to be more susceptible to antibiotics present in the selective media, particularly colistin and polymyxin B, than was C. jejuni. We therefore recommend M2 for the isolation of Campylobacter spp. Finally, the filtration method was found to be easy and cheap; although the sensitivity was low, this method allowed the isolation of new Campylobacter spp. which seem to be associated with diarrhea.

摘要

我们之前描述过的(H. 古森斯、M. 德博克和J. P. 布茨勒,《欧洲临床微生物学杂志》2:389 - 393,1983年)用于从粪便标本中分离空肠弯曲菌和结肠弯曲菌的选择性培养基,其组成为头孢哌酮(15毫克/升)、利福平(10毫克/升)、多粘菌素(10000国际单位/升)和两性霉素B(2毫克/升)(培养基M1),现作如下修改:头孢哌酮(30毫克/升)、利福平(10毫克/升)和两性霉素B(2毫克/升)(培养基M2)。使用培养基M1、培养基M2、一种不含血液的选择性培养基(每升含)活性炭(4克)、硫酸亚铁(0.25克)、丙酮酸钠(0.25克)、酪蛋白水解物(3克)、脱氧胆酸钠(1克)、2号营养肉汤(25克)、琼脂(12克)和头孢哌酮(32毫克)(培养基M3)以及含有甲氧苄啶(10毫克)、利福平(10毫克)、多粘菌素B(5000国际单位)和环己酰亚胺(100毫克)(每升)的普雷斯顿培养基(培养基M4)对从粪便标本中分离弯曲菌属进行了一项比较研究。我们还纳入了一个过滤系统,其中将膜过滤器直接应用于分布在小培养皿中的非选择性无血培养基表面。总共检测了5276份粪便标本:在研究1中,2788份粪便标本在M1和M3上进行检测;在研究2中,2488份粪便标本接种于四种选择性培养基上,并且在这2488份标本中的最后986份与过滤系统平行检测。在研究2中,从126名不同患者中分离出128株弯曲菌菌株;这些菌株分别有85.0%、88.3%、82.5%和66.6%在M1、M2、M3和M4上分离得到。在M1、M2、M3和M4平板上分别有65.4%、70.7%、62.4%和40.3%未发现污染的粪便菌群。此外,发现结肠弯曲菌比空肠弯曲菌对选择性培养基中存在的抗生素,特别是多粘菌素和多粘菌素B更敏感。因此,我们推荐使用M2来分离弯曲菌属。最后,发现过滤方法简便且成本低廉;尽管灵敏度较低,但该方法能够分离出似乎与腹泻相关的新的弯曲菌属菌种。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8d3f/269038/32cd29d2d50b/jcm00101-0177-a.jpg

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