Field Adaptation of an Isothermal Amplification Technique for the Diagnosis of Brucellosis, Contagious Caprine Pleuropneumonia and Peste des petits ruminants in Goats

Loop mediated isothermal amplification (LAMP) is a simple and powerful innovative gene amplification technique; used as a rapid diagnostic tool for early detection and identification of microbial diseases. The whole procedure can be completed in less than one hour under isothermal conditions. Thus, provide a low cost, highly sensitive, highly specific and less laborious alternative to detect infectious diseases. LAMP has all the characteristics required for real-time assays (quantitative) along with the simple operation for easy adaptability to field conditions. LAMP technique amplifies DNA with high efficiency under isothermal conditions without a significant influence of the co-presence of non-target DNA. Its detection limit is a few copies, being comparable to that of polymerase chain reaction. LAMP is easy to perform once the appropriate primers are designed. It requires four to six primers, a DNA polymerase and a regular laboratory water bath or heat block for the completion of the reaction. By combination with reverse transcription, LAMP can amplify RNA sequences with high efficiency. A great advantage of the technique is that gene amplification can be visualized by the naked eye either as turbidity or in the form of a color change when SYBR Green I, a fluorescent dsDNA intercalating dye, is employed. Alternatively, products can be detected by agarose gel electrophoresis. The endemic status of brucellosis, contagious caprine pleuropneumonia and peste des petits ruminants diseases in Oman, continues to devastate the animal wealth sector. For the control and eradication of such diseases; adaptation of the LAMP technique for disease diagnosis under field conditions would be an ideal choice particularly with its low-cost benefits. We would like to establish the first LAMP unit in Oman for the diagnosis of various infectious diseases of veterinary importance. The unit will provide diagnostic capabilities, training and expertise for veterinarians and other personals who are interested in the different applications of the LAMP technology.