Wasfy M, Oyofo B, Elgindy A, Churilla A
Enteric Microbiology Branch, U.S. Naval Medical Research Unit no. 3, Cairo, Egypt.
J Clin Microbiol. 1995 Aug;33(8):2176-8. doi: 10.1128/jcm.33.8.2176-2178.1995.
Transportation of clinical samples and long-term recoverability of pathogens are critical to epidemiological studies, particularly when conditions do not permit immediate processing. This study confirms that Cary-Blair medium (CB) is suitable for the preservation of Salmonella and Shigella isolates for more than 2 weeks at 25, 4, or -70 degrees C. Campylobacter jejuni was not recovered after 2 days of storage in CB at 25 degrees C when an inoculum of 12 x 10(8) cells per ml was used. Lower temperatures supported the recovery of this organism for 6 days. When individual pathogens were preserved with stools in CB and incubated at 25, 4, or -70 degrees C, the Salmonella and Shigella concentrations dropped from 12 x 10(8) cells to 1 x 10(3) or 1 x 10(4) cells per ml within 2 days and then remained stable for the rest of the observation period (15 days). C. jejuni survived preservation with stools for 5 to 9 days. The addition of blood and glycerol to CB improved the recoverability of all enteropathogens, particularly C. jejuni, which was consistently detected for 7 to 9 days at the different preservation temperatures used. When trypticase soy broth-glycerol (freezing medium), with or without blood, was used, there was little or no decrease in the Salmonella and Shigella concentrations during 2 weeks of preservation with stools at -70 degrees C. C. jejuni demonstrated a relatively sustained high concentration in Trypticase soy broth-glycerol with 5% blood. The use of defibrinated, laked sheep blood as a long-term freezing medium supported the recovery of low concentrations of Salmonella and Shigella spp. (10(2) to 10(3) cells per ml) for more than 14 weeks. Recovery of C. jejuni was consistent for 7 weeks when an initial concentration of 10(6) cells per ml present in stools. Laked blood provided a simple, sterile, and inexpensive medium for the preservation of individual isolates and clinical samples.
临床样本的运输以及病原体的长期可恢复性对于流行病学研究至关重要,尤其是在条件不允许立即处理的情况下。本研究证实, Cary - Blair培养基(CB)适用于在25、4或 -70摄氏度下保存沙门氏菌和志贺氏菌分离株超过2周。当使用每毫升12×10⁸个细胞的接种量时,空肠弯曲菌在25摄氏度的CB中储存2天后无法恢复。较低温度下该菌可恢复6天。当将单个病原体与粪便一起保存在CB中并在25、4或 -70摄氏度下孵育时,沙门氏菌和志贺氏菌的浓度在2天内从每毫升12×10⁸个细胞降至1×10³或1×10⁴个细胞,然后在其余观察期(15天)内保持稳定。空肠弯曲菌与粪便一起保存可存活5至9天。向CB中添加血液和甘油可提高所有肠道病原体的可恢复性,特别是空肠弯曲菌,在使用的不同保存温度下,其可连续7至9天被检测到。当使用胰蛋白酶大豆肉汤 - 甘油(冷冻培养基),无论有无血液,在 -70摄氏度下与粪便一起保存2周期间,沙门氏菌和志贺氏菌的浓度几乎没有下降。空肠弯曲菌在含5%血液的胰蛋白酶大豆肉汤 - 甘油中显示出相对持续的高浓度。使用去纤维蛋白的羊血作为长期冷冻培养基可支持低浓度沙门氏菌和志贺氏菌属(每毫升10²至10³个细胞)超过14周的恢复。当粪便中初始浓度为每毫升10⁶个细胞时,空肠弯曲菌的恢复持续7周。羊血为保存单个分离株和临床样本提供了一种简单、无菌且廉价的培养基。