Voss Andreas, Mouton Johan W, van Elzakker Erika P, Hendrix Ron G, Goessens Wil, Kluytmans Jan A, Krabbe Paul F, de Neeling Han J, Sloos Jacobus H, Oztoprak Nefise, Howe Robin A, Walsh Timothy R
Radboud University Nijmegen Medical Centre, Nijmegen University Centre of Infectious Diseases, The Netherlands.
Ann Clin Microbiol Antimicrob. 2007 Sep 24;6:9. doi: 10.1186/1476-0711-6-9.
To determine the true incidence of hGISA/GISA and its consequent clinical impact, methods must be defined that will reliably and reproducibly discriminate these resistant phenotypes from vancomycin susceptible S. aureus (VSSA).
This study assessed and compared the ability of eight Dutch laboratories under blinded conditions to discriminate VSSA from hGISA/GISA phenotypes and the intra- and inter-laboratory reproducibility of agar screening plates and the Etest method. A total of 25 blinded and unique strains (10 VSSA, 9 hGISA and 6 GISA) were categorized by the PAP-AUC method and PFGE typed to eliminate clonal duplication. All strains were deliberately added in quadruplets to evaluate intra-laboratory variability and reproducibility of the methods. Strains were tested using three agar screening methods, Brain Heart Infusion agar (BHI) + 6 microg/ml vancomycin, Mueller Hinton agar (MH) + 5 microg/ml vancomycin and MH + 5 microg/ml teicoplanin) and the Etest macromethod using a 2 McFarland inoculum.
The ability to detect the hGISA/GISA phenotypes varied significantly between methods and phenotypes. BHI vancomycin and MH vancomycin agar screens lacked the ability to detect hGISA. The MH teicoplanin agar screen was more sensitive but still inferior to Etest that had a sensitivity of 98.5% and 99.5%, for hGISA and GISA, respectively. Intra- and inter-laboratory reproducibility varied between methods with poorest performance seen with BHI vancomycin.
This is the first multi-center blinded study to be undertaken evaluating various methods to detect GISA and hGISA. These data showed that the ability of clinical laboratories to detect GISA and hGISA varied considerably, and that screening plates with vancomycin have a poor performance in detecting hGISA.
为了确定异质性耐万古霉素金黄色葡萄球菌(hGISA)/耐万古霉素金黄色葡萄球菌(GISA)的真实发病率及其相应的临床影响,必须定义能够可靠且可重复地将这些耐药表型与万古霉素敏感金黄色葡萄球菌(VSSA)区分开来的方法。
本研究评估并比较了荷兰8个实验室在盲法条件下区分VSSA与hGISA/GISA表型的能力,以及琼脂筛选平板和Etest方法在实验室内和实验室间的可重复性。通过PAP-AUC方法对总共25株盲法且独特的菌株(10株VSSA、9株hGISA和6株GISA)进行分类,并通过脉冲场凝胶电泳(PFGE)分型以消除克隆重复。所有菌株均故意以四联形式添加,以评估方法在实验室内的变异性和可重复性。使用三种琼脂筛选方法[脑心浸液琼脂(BHI)+6μg/ml万古霉素、穆勒-欣顿琼脂(MH)+5μg/ml万古霉素和MH+5μg/ml替考拉宁]以及使用2麦氏浊度接种物的Etest宏方法对菌株进行检测。
不同方法和表型之间检测hGISA/GISA表型的能力差异显著。BHI万古霉素和MH万古霉素琼脂筛选缺乏检测hGISA的能力。MH替考拉宁琼脂筛选更敏感,但仍不如Etest,Etest对hGISA和GISA的敏感性分别为98.5%和99.5%。不同方法在实验室内和实验室间的可重复性各不相同,其中BHI万古霉素的表现最差。
这是第一项评估检测GISA和hGISA的各种方法的多中心盲法研究。这些数据表明,临床实验室检测GISA和hGISA的能力差异很大,且含万古霉素的筛选平板在检测hGISA方面表现不佳。