Development and evaluation of loop-mediated isothermal amplification assay for rapid detection of Capripoxvirus

Abstract

Aim: The present study was undertaken to develop a nucleic acid-based diagnostic assay loop-mediated isothermal amplification assay (LAMP) targeting highly conserved genomic regions of Capripoxvirus (CaPVs) and its comparative evaluation with real-time polymerase chain reaction (PCR). Material and Methods: Lyophilized vaccine strain of sheeppox virus (SPPV) was used for optimization of LAMP assay. The LAMP assay was designed using envelope immunogenic protein (P32) coding gene targeting highly conserved genomic regions of CaPV responsible for causing sheep pox, goat pox, and lumpy skin disease in sheep, goat and cattle respectively. Serial tenfold dilution of SPPV recombinant plasmid DNA was used for a calculating limit of detection. Analytical sensitivity and specificity were performed. Results: The test described is quick (30 min), sensitive and specific for detection of CaPVs. The described assay did not show any cross-reactivity to other related viruses that cause apparently similar clinical signs. It was found to be ten times more sensitive than conventional PCR however, 100 times less sensitive than quantitative PCR (qPCR). LAMP assay results were monitored by color change method using picogreen dye and agarose gel electrophoresis. Conclusion: LAMP assay can be a very good alternative for CaPV detection to other molecular techniques requiring sophisticated equipments.

The Effects of Moderate Physical Exercise on Adult Cognition: A Systematic Review

“…In the articles included in this review (Bue-Estes et al, 2008 ; Tsai et al, 2014 ; Olson et al, 2016 ), reaction time as a cognitive parameter was found to be improved, as seen in Table 4 . This is an important parameter that can help evaluate the speed of information processing (Batra et al, 2015 ). These processes were analyzed for their speed and accuracy in terms of response (van Ede et al, 2012 ), thus evidencing a possible association between moderate physical exercise and improvement of the synaptic network integration involved in this action.…”

Establishment of an accurate and fast detection method using molecular beacons in loop-mediated isothermal amplification assay

“…These characteristics provide results in a fast, efficient, and cost-effective method that theoretically should amplify target sequences with higher specificity than PCR. Since its introduction, LAMP has seen wide interdisciplinary application, for example in detecting microorganisms (e.g., bacteria23, fungi4, and viruses56) diagnosing genetic diseases7, and even early sex determination8.…”

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

“…Other dyes such as malachite green dye (Sriworarat et al 2015;Lucchi et al 2016;Nzelu et al 2016), Picogreen dye (Batra et al 2015), Gelred dye (Wozniakowski et al 2013;Wassermann et al 2014), SYTO fluorescent dye (Liu et al 2013;Watts et al 2014;Mwendwa et al 2017) Zhou et al, 2014). The reaction between free calcein and magnesium ions intensified the fluorescence of LAMP product (ii.)…”