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.
Highlights
Escherichia coli O157:H7 is an enterohemorrhagic strain of the bacterium E. coli and a cause of food-borne illness
Considering the state of art status of Loop Mediated Isothermal Amplification (LAMP) technique that has achieved its advancement for the diagnosis of various viruses worldwide, the current study emphasizes on its application on a bacterial pathogen and can be extended towards parasitic and fungal pathogens in future
LAMP from research lab to clinical diagnosis: it is suggested here that the technologies associated with LAMP can be considered and developed as part of a LAMP platform, rather than developing them as separate entities
Summary
Escherichia coli O157:H7 is an enterohemorrhagic strain of the bacterium E. coli and a cause of food-borne illness. Most illness has been associated with eating undercooked, contaminated ground beef, drinking unpasteurized milk, swimming in or drinking contaminated water and eating contaminated vegetables. Young children and females had an increased risk of Hemolytic Uremic Syndrome (HUS) after Shiga toxin producing E. coli (STEC) O157 infection. With or without HUS, elderly persons had the highest pro-portion of deaths associated with STEC O157 infection. These data support recommendations for aggressive sup-portive care of young children and the elderly early during illness due to STEC O157 (Gould et al, 2009). E. coli O157:H7 was the causative agent of many out-breaks worldwide.
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