Perçin Duygu, Colakoğlu Selcan, Durmaz Süleyman, Ekincioğlu Pınar
Department of Medical Microbiology, Erciyes University Faculty of Medicine, Kayseri, Turkey.
Mikrobiyol Bul. 2012 Oct;46(4):546-52.
Detection of rectal colonization with carbapenem-resistant Klebsiella pneumoniae (CRKP) is the most important step in the infection control protocols in order to prevent infections caused by CRKP which has an increasing incidence all over the world. In this study, it was aimed to compare the detection rate of 2 mg/L ertapenem EMB agar medium with the other methods recommended by various international guidelines. These methods include direct plate method using ertapenem disc, enrichment method in tryptic soy broth containing 2 mg/L ertapenem and the investigation of the predominant betalactamases in the colonized patients. The lowest inoculum detected by different methods was determined by using simulative challenge test prepared for this purpose. The ability to detect CRKP from rectal swabs was evaluated by using the clinical specimens of 801 patients. For all bacteria isolated, carbapenem susceptibility was evaluated by using E-test method, the presence of beta-lactamases was determined by using modified Hodge test (MHT), and the carbapenemase genes were investigated by using multiplex polymerase chain reaction (PCR). The lowest inoculum detected by ertapenem-EMB agar was 50 CFU/mL whereas the lowest inocula were 1 x 105 and 1 x 103, respectively by tryptic soy broth with ertapenem and direct plate method. No resistance gene were identified by PCR in 13 (39.4%) of 33 isolates, whereas blaOXA-48 was detected in 19 (95%) and blaIMP in 1 (5%) of 20 positive isolates. All of the positive strains were resistant to imipenem and ertapenem, while 2 (10%) strains were found to be susceptible to doripenem and meropenem. While MHT was negative in all strains which were negative for resistance genes, all resistance gene positive strains except one blaOXA-48 strain that was also sensitive to doripenem and meropenem, were found to be positive with MHT. According to the results of PCR, the sensitivities of the three methods were found to be 80%. The specificities, positive and negative predictive values were found to be 15.4%, 59% and 33.3% for ertapenem-EMB agar, 23%, 61.5% and 42.9% for broth with ertapenem and 61.5%, 76% and 66.6% for direct plate method, respectively. Average labor time of the methods (isolation + identification + sensitivity + MHT) was determined as 48 hours for ertapenem-EMB agar, whereas it was 96 hours for the other methods. In conclusion, since ertapenem- EMB agar method is a sensitive and rapid method, it can be used safely for the preliminary detection of CRKP without increasing the workload of the laboratory.
检测耐碳青霉烯类肺炎克雷伯菌(CRKP)的直肠定植是感染控制方案中预防由CRKP引起感染的最重要步骤,CRKP在全球的发病率呈上升趋势。本研究旨在比较2mg/L厄他培南伊红美蓝琼脂培养基与各种国际指南推荐的其他方法的检测率。这些方法包括使用厄他培南纸片的直接平板法、在含2mg/L厄他培南的胰蛋白胨大豆肉汤中的增菌法以及对定植患者中主要β-内酰胺酶的检测。通过为此目的制备的模拟挑战试验确定不同方法检测到的最低接种量。使用801例患者的临床标本评估从直肠拭子中检测CRKP的能力。对于所有分离出的细菌,使用E-test法评估碳青霉烯类药物敏感性,使用改良 Hodge试验(MHT)确定β-内酰胺酶的存在,并使用多重聚合酶链反应(PCR)研究碳青霉烯酶基因。厄他培南-伊红美蓝琼脂检测到的最低接种量为50CFU/mL,而含厄他培南的胰蛋白胨大豆肉汤和直接平板法的最低接种量分别为1×10⁵和1×10³。在33株分离株中的13株(39.4%)中,PCR未鉴定出耐药基因,而在20株阳性分离株中的19株(95%)中检测到blaOXA - 48,1株(5%)中检测到blaIMP。所有阳性菌株对亚胺培南和厄他培南耐药,而2株(10%)菌株对多利培南和美罗培南敏感。在所有耐药基因阴性的菌株中MHT均为阴性,除1株对多利培南和美罗培南也敏感的blaOXA - 48菌株外,所有耐药基因阳性菌株的MHT均为阳性。根据PCR结果,三种方法的敏感性均为80%。厄他培南-伊红美蓝琼脂的特异性、阳性和阴性预测值分别为15.4%、59%和33.3%,含厄他培南肉汤的分别为23%、61.5%和42.9%,直接平板法的分别为61.5%、76%和66.6%。这些方法的平均 labor时间(分离+鉴定+敏感性+MHT),厄他培南-伊红美蓝琼脂法确定为48小时,而其他方法为96小时。总之,由于厄他培南-伊红美蓝琼脂法是一种敏感且快速的方法,它可以安全地用于CRKP的初步检测,而不会增加实验室的工作量。 (注:原文中“labor time”表述有误,推测可能是“laboratory time”,翻译时按“实验室时间”理解,但不确定是否正确,按原文照译)