Simple, Powerful, and Smart: Using LAMP for Low Cost Screening of Multiple Waterborne Pathogens
Grégoire Seyrig, Farhan Ahmad, Robert D. Stedtfeld, Dieter M. Tourlousse and Syed A. Hashsham
from: Environmental Microbiology: Current Technology and Water Applications (Edited by: Keya Sen and Nicholas J. Ashbolt). Caister Academic Press, U.K. (2011)
A vast array of low cost, simple, rugged, and rapid molecular approaches are emerging for the detection of indicators and pathogens, along with the collection of relevant genotypic information. This chapter focuses on loop-mediated isothermal amplification (LAMP), a relatively new DNA amplification technique, which due to its simplicity, ruggedness, and low cost could provide major advantages to the water industry. In LAMP, the target sequence is amplified at a constant temperature using either two or three sets of primers and a polymerase with high strand displacement activity. Due to the specific nature of the action of these primers, the amount of DNA produced in LAMP is considerably higher than PCR based amplification. The corresponding release of pyrophosphate results in visible turbidity due to precipitation, which allows easy visualization by the naked eye, especially for larger reaction volumes or via simple detection approaches for smaller volumes. The reaction can be followed in real-time either by measuring the turbidity or the signals from DNA produced via fluorescent dyes that intercalate or directly label the DNA, and in turn can be correlated to the number of copies initially present. Hence, LAMP can also be quantitative. While LAMP is already the method of choice in organizations engaged in combating infectious diseases such as tuberculosis, malaria, and sleeping sickness in developing regions, it has yet to be extensively validated for commonly known waterborne pathogens read more ...