Whole animal automated platform for drug discovery against multi-drug resistant Staphylococcus aureus.
Whole animal automated platform for drug discovery against multi-drug resistant Staphylococcus aureus.
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DOI：
10.1371/journal.pone.0089189
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Mylonakis E
中科院分区：
文献类型：
--
作者：
Rajamuthiah R;Fuchs BB;Jayamani E;Kim Y;Larkins-Ford J;Conery A;Ausubel FM;Mylonakis E
金黄色葡萄球菌是美国医院获得性感染的主要原因，也是模式线虫秀丽隐杆线虫的致病性。梭elegans-S.金黄色葡萄球菌感染模型先前在固体琼脂平板上进行，其中食菌性C.秀丽线虫以S.金黄色。然而，基于琼脂的测定不适合于抗菌化合物的大规模筛选。我们已经开发了一种高通量液体筛选检测方法，该方法使用机器人仪器来分配精确量的甲氧西林耐药S。金黄色葡萄球菌（MRSA）和蠕虫在384孔测定板中进行，然后进行自动显微镜和图像分析。在液体试验的验证中，发现MRSA细胞壁缺陷型突变体MW 2 ΔtarO在基于琼脂的试验中因杀灭而减毒，在液体试验中毒性较低。利用Z '因子始终大于0.5的这种稳健测定来筛选由640种具有良好表征的生物活性的小分子组成的Biomol 4化合物文库。作为原理的证明，库中存在的30种临床使用的抗生素中有27种赋予增加的C。elegans存活，并被鉴定为筛选中的命中。令人惊讶的是，抗蠕虫药物氯氰碘柳胺也被确定为屏幕上的热门。在进一步的研究中，我们证实了氯氰碘柳胺对万古霉素耐药葡萄球菌的抗葡萄球菌活性。金黄色葡萄球菌分离株和其他革兰氏阳性菌。液体C。elegans - S.本文所述的金黄色葡萄球菌测定允许筛选对宿主无毒的抗葡萄球菌化合物。
Staphylococcus aureus, the leading cause of hospital-acquired infections in the United States, is also pathogenic to the model nematode Caenorhabditis elegans. The C. elegans-S. aureus infection model was previously carried out on solid agar plates where the bacteriovorous C. elegans feeds on a lawn of S. aureus. However, agar-based assays are not amenable to large scale screens for antibacterial compounds. We have developed a high throughput liquid screening assay that uses robotic instrumentation to dispense a precise amount of methicillin resistant S. aureus (MRSA) and worms in 384-well assay plates, followed by automated microscopy and image analysis. In validation of the liquid assay, an MRSA cell wall defective mutant, MW2ΔtarO, which is attenuated for killing in the agar-based assay, was found to be less virulent in the liquid assay. This robust assay with a Z’-factor consistently greater than 0.5 was utilized to screen the Biomol 4 compound library consisting of 640 small molecules with well characterized bioactivities. As proof of principle, 27 of the 30 clinically used antibiotics present in the library conferred increased C. elegans survival and were identified as hits in the screen. Surprisingly, the antihelminthic drug closantel was also identified as a hit in the screen. In further studies, we confirmed the anti-staphylococcal activity of closantel against vancomycin-resistant S. aureus isolates and other Gram-positive bacteria. The liquid C. elegans – S. aureus assay described here allows screening for anti-staphylococcal compounds that are not toxic to the host.
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DOI：
10.1073/pnas.1209126109
发表时间：
2012-11-13
影响因子：
11.1
作者：
Brown, Stephanie;Xia, Guoqing;Walker, Suzanne
通讯作者：
Walker, Suzanne
影响因子：
5.2
作者：
Day, Shandra R.;Moore, Christopher M.;Sifri, Costi D.
通讯作者：
Sifri, Costi D.
影响因子：
5.2
作者：
Gottlieb, GS;Fowler, VG;Corey, GR
通讯作者：
Corey, GR
影响因子：
4
作者：
Campbell, Jennifer;Singh, Atul K.;Maria, John P. Santa, Jr.;Kim, Younghoon;Brown, Stephanie;Swoboda, Jonathan G.;Mylonakis, Eleftherios;Wilkinson, Brian J.;Walker, Suzanne
通讯作者：
Walker, Suzanne
影响因子：
14.2
作者：
Benfield, T.;Espersen, F.;Skinhoj, P.
通讯作者：
Skinhoj, P.