Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence.
Development and Validation of a Microtiter Plate-Based Assay for Determination of Bacteriophage Host Range and Virulence.
复制标题
DOI：
10.3390/v10040189
复制
发表时间：
2018-04-12
期刊：
影响因子：
--
通讯作者：
Gill JJ
中科院分区：
文献类型：
--
作者：
Xie Y;Wahab L;Gill JJ
噬菌体是细菌的天敌，它已重新成为对抗抗生素耐药细菌的有吸引力的替代品。噬菌体在物种和菌株水平上具有高度特异性，噬菌体宿主范围的测量在利用噬菌体作为抗菌剂方面起着重要作用。确定噬菌体宿主范围的最常用方法是在软琼脂覆盖物上发现噬菌体裂解物并观察噬菌斑形成。在本研究中，采用 96 孔微量滴定板形式开发了一种基于液体培养的测定法，以测量 15 种沙门氏菌噬菌体集合对代表 11 种血清型的 20 种沙门氏菌菌株的噬菌体宿主范围和毒力。该方法与传统的点法进行了比较。两种方法得出的大多数宿主范围结果是一致的，包括细菌菌株对噬菌体不敏感的情况。与其他方法相比，每种方法在 19/300 (6%) 的测量噬菌体-宿主组合中产生假阴性结果。点法往往比微量滴定法显示出更高的噬菌体敏感性，尽管直接比较两种方法之间的响应幅度很困难，因为它们的作用机制不同。微量滴定板测定能够以高通量格式提供更高分辨率的噬菌体宿主范围和毒力数据。
Bacteriophages, which are the natural predators of bacteria, have re-emerged as an attractive alternative to combat antibiotic resistant bacteria. Phages are highly specific at the species and strain level and measurement of the phage host range plays an important role in utilizing the phage as antimicrobials. The most common method for phage host range determination has been to spot phage lysates on soft agar overlays and observe plaque formation. In this study, a liquid culture-based assay was developed in a 96-well microtiter plate format to measure the phage host range and virulence for a collection of 15 Salmonella phages against a panel of 20 Salmonella strains representing 11 serovars. This method was compared to a traditional spot method. The majority of the host range results from two methods were in agreement including in cases where a bacterial strain was insensitive to the phage. Each method produced a false-negative result in 19/300 (6%) of the measured phage-host combinations when compared to the other method. The spot method tended to indicate greater phage sensitivity than the microtiter assay even though direct comparisons of the response magnitude between the two methods is difficult since they operate on different mechanisms. The microtiter plate assay was able to provide data on both the phage host range and virulence in greater resolution in a high-throughput format.
登录
查看更多内容
影响因子：
5.2
作者：
Bull JJ;Gill JJ
通讯作者：
Gill JJ
影响因子：
5.2
作者：
Bryan D;El-Shibiny A;Hobbs Z;Porter J;Kutter EM
通讯作者：
Kutter EM
影响因子：
5.2
作者：
Andrews, JM
通讯作者：
Andrews, JM
影响因子：
4.8
作者：
El-Arabi TF;Griffiths MW;She YM;Villegas A;Lingohr EJ;Kropinski AM
通讯作者：
Kropinski AM
影响因子：
3.7
作者：
Ahmed SA;Awosika J;Baldwin C;Bishop-Lilly KA;Biswas B;Broomall S;Chain PS;Chertkov O;Chokoshvili O;Coyne S;Davenport K;Detter JC;Dorman W;Erkkila TH;Folster JP;Frey KG;George M;Gleasner C;Henry M;Hill KK;Hubbard K;Insalaco J;Johnson S;Kitzmiller A;Krepps M;Lo CC;Luu T;McNew LA;Minogue T;Munk CA;Osborne B;Patel M;Reitenga KG;Rosenzweig CN;Shea A;Shen X;Strockbine N;Tarr C;Teshima H;van Gieson E;Verratti K;Wolcott M;Xie G;Sozhamannan S;Gibbons HS;Threat Characterization Consortium
通讯作者：
Threat Characterization Consortium