Determination of Chikungunya Virus RNA by Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) Assay

Abstract

As a matter of fact, infectious diseases hamper everyone’s life and produce a lifelong threat to everyone neutral of age, sex, lifestyle and socio-economic status. Nowadays, come into the sight of Chikungunya viral (CHIKV) infection injured many Asian and African countries, also deliberated threat in rising countries and also low socio-economic countries. CHIKV is a positive-sense, enveloped single-stranded, RNA virus belonging to the genus Alphavirus, family Togaviridae. As the Dengue & Chikungunya viruses are spread simultaneously at the same time, so it is tough to identify them. In our resource-limited countries, swift detection of CHIKV by RT-LAMP is the simplest molecular technique in low-equipment settings without the use of any expensive decoration. Heat-treated centrifuged and uncentrifuged samples were used in this study and they showed the same result (100%). Different instruments like heat block, water bath, conventional thermal cycler & real-time thermal cycler were used to amplify the CHIKV RNA and they indicated that 100% samples were identified by all four instruments. The amplified products were visualized by turbidity test, color change by HNB, step-ladder band pattern in agarose gel electrophoresis and amplification curve in real-time thermal cycler naked eye, here the results also showed 100% samples were determined by all visualized methods. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a recent technique for amplifying RNA under limited temperature, with high tangibility, quickness and competency. To identify the CHIKV RNA Reverse transcription loop-mediated isothermal amplification was fabricated and validated, and the results were also compared with reverse transcription polymerase chain reaction (RT-PCR). The sensitivity was 95.71% and specificity was 100%, these results indicate that RT-LAMP is a feasible method for quick detection of CHIKV RNA.