Evaluation of Two Lyophilized Molecular Assays to Rapidly Detect Foot-and-Mouth Disease Virus Directly from Clinical Samples in Field Settings

• DOI: 10.1111/tbed.12451
• Publication Date: 2015-11-30T10:41:29Z
• Authors: E. L. A. Howson; B. Armson; M. Madi; C. J. Kasanga; S. Kandusi; R. Sallu; E. Chepkwony; A. Siddle; P. Martin; J. Wood; V. Mioulet; D. P. King; T. Lembo; S. Cleaveland; V. L. Fowler
• Keywords: Standard operating procedure

Accurate, timely diagnosis is essential for the control, monitoring and eradication of foot-and-mouth disease (FMD). Clinical samples from suspect cases are normally tested at reference laboratories. However, transport to these centralized facilities can be a lengthy process that impose delays on critical decision making. These concerns have motivated work evaluate simple-to-use technologies, including molecular-based diagnostic platforms, deployed closer FMD. In this context, FMD virus (FMDV)-specific reverse transcription loop-mediated isothermal amplification (RT-LAMP) real-time RT-PCR (rRT-PCR) assays, compatible with simple sample preparation methods in situ visualization, been developed which share equivalent analytical sensitivity laboratory-based rRT-PCR. lack robust 'ready-to-use kits' utilize stabilized reagents limits deployment tests into field settings. To address gap, study describes performance lyophilized rRT-PCR RT-LAMP assays detect FMDV. Both use fluorescence monitor real-time, end point detection could also achieved using molecular lateral flow devices. Lyophilization did not adversely affect assays. Importantly, when were challenging laboratory settings within East Africa they proved reliable their ability FMDV range clinical acutely infected as well convalescent cattle. data support highly sensitive rapid