Abstract

Loop Mediated Isothermal Amplification (LAMP) assay could be a useful adjunct diagnostic assay along with the conventional methods that would preclude the requirement of continuous maintenance of pure cultures. Moreover, LAMP assay is simple, rapid, specific and sensitive for the detection of pathogens. Having developed and validated LAMP method for the detection of an isolated pathogen, Escherichia coliO157:H7, an attempt was made to progress the LAMP platform to realistic point of care for resource-poor endemic areas. Reaction time of the LAMP method was only 1 h and also, the amplification products of O157, which had the corresponding target genes, turned green by visual inspection when added with Calcein/Sybr green. However, sample preparation and lyophilized master mix preparation before LAMP assay as well as developing a closed detection system for detection of LAMP amplified products remained a quest. Hence, the current study was conducted to develop a lyophilized LAMP master mix for easy platform to take LAMP to realistic point of care and Dot-Elisa based ‘lateral flow dipstick’ that could ease the detection of LAMP products in a closed environment. Sample preparation is another associated technology that is yet-to-be developed.

Key words: Loop mediated isothermal amplification assay, polymerase chain reaction, lyophilized master mix, and closed amplification system.