Development of a loop-mediated isothermal amplification method for rapid mass-screening of sand flies for Leishmania infection

Abstract

Entomological monitoring of Leishmania infection in leishmaniasis endemic areas offers epidemiologic advantages for predicting the risk and expansion of the disease, as well as evaluation of the effectiveness of control programs. In this study, we developed a highly sensitive loop-mediated isothermal amplification (LAMP) method for the mass screening of sand flies for Leishmania infection based on the 18S rRNA gene. The LAMP technique could detect 0.01 parasites, which was more sensitive than classical PCR. The method was robust and could amplify the target DNA within 1 hr from a crude sand fly template without DNA purification. Amplicon

Simple and Rapid Visual Detection Methods of Orf Virus by B2L Gene based Loop-Mediated Isothermal Amplification Assay

“…The difficulty associated with detection methods of LAMP in field laboratory settings will prevent its applicability. But, low cost, ease of visualization without need of any instruments, and plasticity of the metal ion indicators namely HNB (Goto et al, 2009) and malachite green (Nzelu et al, 2014;Nathan et al, 2015) could be handful to achieve a point of care diagnostic field applicability of this isothermal amplification assay. Further, visual observation of LAMP reaction by colorimetric detection using SYBR green I and HNB dyes will reduce the time taken for diagnosis in field conditions and avoid handling high cost equipment and unsafe reagents associated with post LAMP amplification analysis (Parida et al, 2008).…”

Development of a loop-mediated isothermal amplification targeting a gene within the pyruvate dehydrogenase complex, the pdhA gene, for rapid detection of Mycoplasma gallisepticum

“…20 Therefore, in order to detect M. gallisepticum infection with limited resources, the development of a cost-efficient, simple, and rapid detection method is required.…”

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

“…The study showed that LAMP was efficient to detect 10 1 copies of DNA. They concluded that the newly developed LAMP was helpful when leishmania parasite density is very low, especially during the early infection stage (Nzelu et al 2014(Nzelu et al , 2016. There was a number of other reported significant practices of LAMP including Trichomonas viganalis (Reyes et al 2014), Necator americanus (Mugambi et al 2015) and Strongyloides stercoralis (Watts et al 2014), and it was shown that LAMP is at least 1000 times more sensitive than the conventional PCR.…”

“…A research was conducted by Nzelu et al (2014) to detect Leishmania. In the study, they developed LAMP and tested it with 122 suspected clinical samples.…”