e., at the sampling site) or at border phytosanitary controls, places where complex facilities may not be available. Loop-mediated isothermal amplification Etomoxir purchase (LAMP) is a novel DNA amplification technique that amplifies DNA with high specificity, efficiency and rapidity under isothermal conditions [17]. LAMP is based on the principle of autocycling strand displacement DNA synthesis performed by the Bst DNA polymerase, for the detection of a specific DNA sequence [17]. The technique uses four to six primers that recognize six to eight regions of the target DNA and provides very high specificity [17, 18]. Amplification can be carried out in a simple and inexpensive device like

a water bath at temperatures between 60 to 65°C. LAMP produces large amounts of DNA [17] and shows high tolerance to biological contaminants [19], thereby simplifying sample preparation. Although LAMP products can be selleck kinase inhibitor detected by gel electrophoresis, this procedure reduces the suitability for field applications. As mentioned above, a LAMP methodology for the detection of Las has been previously reported [11]. That work focused on the detection of the DNA sequence of the tufB-secE-nusG-rplKAJL-rpoB gene cluster present in the microorganism. The analysis

of the amplification products was done by gel electrophoresis, or dot-blotting of the amplification products on a nylon membrane followed by staining with Mupid Blue, methods that are not compatible with field applications. DMXAA cell line On our study, we target a hypothetical protein-coding sequence present in the genome of Las for the detection of this pathogen. To overcome the limitations associated with the gel electrophoresis, we coupled the LAMP amplification with a Lateral

Flow Dipstick (LFD), which permits an accurate and straightforward detection of LAMP amplicons, eliminating the need of complex equipment and data analysis [20, 21]. By using both LAMP and LFD technologies, this work describes the development of a new molecular diagnostic Florfenicol tool for the detection of Las. Results and discussion In order to develop a successful HLB management strategy, methods for rapid detection of pathogens in the field are required. Such detection would allow early diagnosis of an infection focus before its spread. LAMP provides an ideal alternative for detection, as it requires a single incubation temperature and obviates the need for expensive thermal cyclers [17]. The combination of this isothermal DNA amplification technique with LFD devices has proven to be robust and successful in field-capable molecular diagnostics [20–22]. The recent sequencing of Las genome has uncovered new DNA sequences that can be used for pathogen detection through DNA amplification technologies [23]. Using an “in silico” approach, we found a hypothetical protein coding sequence, CLIBASIA_05175 [GenBank: ACT57606.1], which was predicted to be highly specific for Las.