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[采用凝胶扩散法优化葡萄球菌中耐甲氧西林检测的氯化钠浓度选择]

[Choice of the NaCl concentration for optimizing the detection of methicillin resistance in Staphylococcus using the gel diffusion method].

作者信息

Bemer-Melchior P, Drugeon H B

机构信息

Laboratoire de bactériologie, hôpital Laënnec, CHU Nantes, boulevard J. Monod, 44093 Saint-Herblain cedex 1, Nantes, France.

出版信息

Pathol Biol (Paris). 2001 Apr;49(3):216-21. doi: 10.1016/s0369-8114(01)00131-6.

Abstract

The detection of oxacillin resistance is evaluated by the disk diffusion method in a collection of 374 Staphylococcus sp strains. The disk diffusion assay is performed with 5-microgram oxacillin disk and a 10(8) CFU/mL inoculum on Mueller-Hinton agar plates supplemented with 2 or 5% NaCl and incubated at 37 degrees C for 24 h. Strains are considered resistant in accordance to the French recommendations (any growth around the disk is observed). The detection of mecA gene is performed by PCR almost for resistant and discordant strains. Results are concordant for 246 of 256 Staphylococcus aureus strains (182 susceptible and 64 resistant strains) and for 105 of 118 S. epidermidis isolates tested (37 susceptible and 68 resistant strains). Six mecA-negative strains (3 S. aureus and 3 S. epidermidis) give false resistant results on agar with 2 and 5% NaCl. Seventeen isolates are discordant on 5% NaCl: 7 mecA-negative S. aureus strains are susceptible on 2% NaCl agar but resistant at 5% (3% false-positive results), 10 mecA-positive S. epidermidis strains are resistant on 2% NaCl agar but susceptible at 5% NaCl (5% false-negative results). The detection of meticillin resistance is improved on agar supplemented with 2% NaCl.

摘要

采用纸片扩散法,对374株葡萄球菌属菌株进行苯唑西林耐药性检测。在补充有2%或5%氯化钠的穆勒-欣顿琼脂平板上,使用含5微克苯唑西林的纸片和10(8) CFU/mL接种物进行纸片扩散试验,并在37℃孵育24小时。根据法国的建议(观察到纸片周围有任何生长),菌株被判定为耐药。几乎对所有耐药和结果不一致的菌株通过聚合酶链反应(PCR)检测mecA基因。对256株金黄色葡萄球菌菌株中的246株(182株敏感菌株和64株耐药菌株)以及118株表皮葡萄球菌分离株中的105株(37株敏感菌株和68株耐药菌株)检测结果一致。6株mecA阴性菌株(3株金黄色葡萄球菌和3株表皮葡萄球菌)在含2%和5%氯化钠的琼脂上呈现假耐药结果。17株菌株在5%氯化钠条件下结果不一致:7株mecA阴性金黄色葡萄球菌菌株在2%氯化钠琼脂上敏感,但在5%氯化钠时耐药(3%假阳性结果),10株mecA阳性表皮葡萄球菌菌株在2%氯化钠琼脂上耐药,但在5%氯化钠时敏感(5%假阴性结果)。在补充有2%氯化钠的琼脂上,甲氧西林耐药性的检测得到了改善。

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