Yamane N, Miyagawa S, Nakasone I, Sakamoto F, Tosaka M
Department of Laboratory Medicine, Faculty of Medicine, University of the Ryukyus, Okinawa.
Rinsho Byori. 1997 Apr;45(4):381-90.
The emergence and dissemination of vancomycin-resistant enterococci (VRE) emphasizes the need for laboratories to be able to correctly detect them. The study described was conducted to evaluate the test methods presently available in Japan to discriminate between the isolates of VRE and those susceptible (VSE). Among the phenotypic test methods evaluated, an agar screening method which utilized 8 micrograms per ml of vancomycin in Mueller-Hinton agar plate appeared to have a sufficient accuracy. When 23 isolates of vanA positive, 31 of vanB positive, 4 of both positive and 60 of both negative were tested, the sensitivity and specificity to detect VRE were estimated to be 98.3% and 100%, respectively. Also, all the VRE isolates were interpreted as being resistant or intermediate by the E test recently approved in Japan, when the results were read after 48 hr-incubation. Whereas, two disk diffusion tests, Showa disk and NCCLS-based Sensi-disc, were evaluated, but both methods failed to discriminate between VRE and VSE, in particular, between the isolates with vanB positive and negative. The automated system, Vitek GPS-TA produced high frequencies of very major errors; 8.7% for vanA positives and 58% for vanB positives. A total of 1,214 enterococcal isolates from multisite laboratories in Japan, comprising 7 different species, were first tested onto agar screening test plates, but none of isolates represented phenotypic vancomycin resistance. With these results, it can be recommended to detect VRE in clinical microbiology laboratories as follows: First, all the enterococcal isolates will be tested onto the agar screening plates or by the E test. Then, if the isolate is interpreted as being resistant or intermediate, the laboratory should confirm whether it is positive for vanA or vanB by polymerase chain reaction (PCR) specified.
耐万古霉素肠球菌(VRE)的出现与传播凸显了实验室能够正确检测它们的必要性。所描述的这项研究旨在评估日本目前可用于区分VRE分离株与敏感株(VSE)的检测方法。在所评估的表型检测方法中,在穆勒 - 欣顿琼脂平板中使用每毫升8微克万古霉素的琼脂筛选方法似乎具有足够的准确性。对23株vanA阳性、31株vanB阳性、4株双阳性和60株双阴性分离株进行检测时,检测VRE的敏感性和特异性估计分别为98.3%和100%。此外,当在48小时培养后读取结果时,所有VRE分离株根据日本最近批准的E试验被判定为耐药或中介。然而,对两种纸片扩散试验,即昭和纸片和基于美国国家临床实验室标准委员会(NCCLS)的敏感纸片进行了评估,但两种方法均无法区分VRE和VSE,特别是无法区分vanB阳性和阴性的分离株。自动化系统Vitek GPS - TA产生的极重大错误频率很高;vanA阳性为8.7%,vanB阳性为58%。首先对来自日本多个实验室的总共1214株肠球菌分离株(包括7个不同菌种)在琼脂筛选试验平板上进行检测,但没有分离株表现出表型万古霉素耐药性。基于这些结果,建议临床微生物实验室按以下方法检测VRE:首先,所有肠球菌分离株都应在琼脂筛选平板上或通过E试验进行检测。然后,如果分离株被判定为耐药或中介,实验室应通过指定的聚合酶链反应(PCR)确认其vanA或vanB是否为阳性。