Evaluation of a cost effective broth and selective agar combination for the detection of MRSA and Staphylococcus aureus from surveillance specimens using regular workflow.

Edward L Chan, Diane Seales
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Abstract

Objective: To evaluate the use of selective agar and broth combination in a regular laboratory daily workflow.

Design: Swabs from 173 surveillance specimens were inoculated onto half of the Bio-Rad MRSASelect (M), SaSelect (S) and Sheep Blood agars (SBA) and the swab placed in the LIM broth. After overnight incubation, 10 microL of the LIM broth was inoculated onto the other half of the three agars and incubated overnight. All the and examined worked after agars were up approximate 14-18 hours of incubation for day one and two according to the regular workflow of the laboratory, without incubating for the full 24 hours for each incubation day. M agar and SBA were evaluated for Methicillin-Resistant Staphylococcus aureus (MRSA), while the S agar was evaluated for Staphylococcus aureus (SA) based on typical colony morphology development. Colonies on the SBA were picked and processed for definitive identification and cefoxitin susceptibility result.

Setting: Trinity Medical Center, a community hospital with network hospitals

Patients: Patient admitted to the hospital submitted swab for surveillance culture

Results: There were a total of 29 MRSA isolated in the study. On day one, both M agar and SBA detected 14 MRSA (48.3%) and on day two, M agar detected 10 (82.7%), while SBA detected 8 (75.8%) additional MRSA. LIM broth added 5 more MRSA to both agars on day 2, to give M agar a total of 29 (100%) and SBA agar a total of 27 (93.1%) of MRSA from the 173 specimens. There were a total of 62 SA isolated. Both the S agar and SBA isolated 34 (54.8%) on day one and 15 more (79%) on day two. The LIM broth added an additional 13 SA for both agars on day two.

Conclusion: Using half of the agar plate for the initial swab and the other half for the broth creates an economic strategy for the detection of MRSA using the M agar and SA using the S agar. Both the M and S agars provided excellent identification and recovery of MRSA or SA based on color and colony morphology unless the colony was too young for color development. The color morphology from the M and S agars was distinguishable overnight after being subcultured from LIM broth. Working up the specimen according to the workflow of the laboratory without having to wait for each plate to incubate a full 24 hours, can still detect all the targeted organisms within 2 workdays using this cost effective strategy.

利用常规工作流程对监测标本中MRSA和金黄色葡萄球菌的成本有效的肉汤和选择性琼脂组合进行评估。
目的:评价选择性琼脂和肉汤组合在常规实验室日常工作中的应用。设计:将173份监测标本的拭子接种于Bio-Rad MRSASelect (M)、SaSelect (S)和羊血琼脂(SBA)的一半上,并将拭子置于LIM肉液中。孵育过夜后,将10 microL的LIM肉汤接种到另外一半琼脂上孵育过夜。根据实验室的常规工作流程,在第一天和第二天进行大约14-18小时的孵育后,所有被检查的动物都进行了大约14-18小时的孵育,而每个孵育日都没有孵育整整24小时。对M琼脂和SBA进行耐甲氧西林金黄色葡萄球菌(MRSA)检测,对S琼脂进行典型菌落形态发育金黄色葡萄球菌(SA)检测。采集SBA上的菌落进行鉴定和头孢西丁敏感性试验。地点:三位一体医疗中心,一个拥有网络医院的社区医院。患者:入院患者提交拭子进行监测培养。结果:研究中共分离出29株MRSA。在第一天,M琼脂和SBA检测到14例MRSA(48.3%),第二天,M琼脂检测到10例(82.7%),SBA检测到8例(75.8%)。第2天,两种琼脂各添加5个MRSA,使173份标本中M琼脂共29个(100%),SBA琼脂共27个(93.1%)。共分离到62株SA。S琼脂和SBA在第一天分离出34株(54.8%),第二天分离出15株(79%)。LIM肉汤在第二天为两种琼脂添加了额外的13 SA。结论:将一半琼脂板用于初始拭子,另一半用于肉汤,为使用M琼脂检测MRSA和使用S琼脂检测SA创造了一种经济策略。M和S琼脂都能很好地识别和恢复MRSA或基于颜色和菌落形态的SA,除非菌落太年轻而不能发育颜色。M和S琼脂的颜色形态在LIM肉汤中继代培养过夜后可区分。根据实验室的工作流程制作标本,而不必等待每个培养皿孵育整整24小时,使用这种成本效益高的策略,仍然可以在2个工作日内检测到所有目标生物。
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