Loop mediated isothermal amplification: An innovative gene amplification technique for animal diseases

Abstract

India being a developing country mainly depends on livestock sector for its economy. However, nowadays, there is emergence and reemergence of more transboundary animal diseases. The existing diagnostic techniques are not so quick and with less specificity. To reduce the economy loss, there should be a development of rapid, reliable, robust diagnostic technique, which can work with high degree of sensitivity and specificity. Loop mediated isothermal amplification assay is a rapid gene amplification technique that amplifies nucleic acid under an isothermal condition with a set of designed primers spanning eight distinct sequences of the target. This assay can be used as an emerging powerful, innovative gene amplification diagnostic tool against various pathogens of livestock diseases. This review is to highlight the basic concept and methodology of this assay in livestock disease.

Development of a real time reverse transcription loop-mediated isothermal amplification method (RT-LAMP) for detection of a novel swine acute diarrhea syndrome coronavirus (SADS-CoV)

“…LAMP technology is currently used for the nucleic acid detection of a number of animal pathogens (Niessen et al, 2013;Sahoo et al, 2016). The assay offers multiple advantages compared to conventional PCR including a shorter turnaround time (30-60 min) and a sensitivity 1-2 orders of magnitude higher (Huang et al, 2016;Yu et al, 2015).…”

Loop-mediated isothermal amplification (LAMP): a versatile technique for detection of micro-organisms

“…Calcein dye, a metal ion binding fluorophore, which was added before isothermal incubation will form an insoluble salt complex, manganese-pyrophosphate (manganese ion, Mn 2+ from calcein whereas pyrophosphate is the byproduct of LAMP reaction), thus making the reaction fluorescent. Subsequently, the fluorescence is further intensified when free calcein binds to magnesium ions (Mg 2+ from LAMP reaction mixture) and could be easily be observed under UV light (365 nm; Sahoo et al 2016). The reaction between Mg 2+ and calcein also causes the colour changes from orange to green, which could be observed via naked eyes under natural light.…”

“…This technique is inapplicable for cloning or other molecular biology purposes since the final product is a large DNA chain, making it to be less versatile when compared to PCR (Sahoo et al 2016). Furthermore, the multiplexing approaches of LAMP is still considered less successful than PCR as they increase the difficulty of the experimental design and procedures .…”

Detection of HCV genotypes 1b and 2a by a reverse transcription loop‐mediated isothermal amplification assay

“…Loop‐mediated isothermal amplification (LAMP) is a newly nucleic acid‐based amplification method that combines high efficiency, cost‐saving, and visualized product reading . These advantages are attributed to four specific LAMP primers and Bacillus stearothermophilus (Bst) DNA polymerase, which can amplify the target DNA and replace the synthesized DNA strand in the meantime, thus allowing the four specific LAMP primers to complete the reaction more quickly and precisely.…”

“…Loop-mediated isothermal amplification (LAMP) is a newly nucleic acid-based amplification method that combines high efficiency, costsaving, and visualized product reading. [28][29][30] These advantages are attributed to four specific LAMP primers and Bacillus stearothermophilus (Bst) DNA polymerase, which can amplify the target DNA and replace the synthesized DNA strand in the meantime, thus allowing the four specific LAMP primers to complete the reaction more quickly and precisely. In addition to its high efficiency, the LAMP method has two major advantages over conventional polymerase chain reaction (PCR) techniques: firstly, it can synthesize DNA products using a simple device such as water bath or heat block without the need of expensive thermocycler machine; and secondly, results can be assessed visually after adding fluorescent dye to the reaction tubes without the need for a special UV illumination machine.…”