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

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Advances in Microbiology 2022年第7期12卷 398-414页

作者： Barnali Sen Sabrina Afrin Sharmin Zahan Saif Ullah Munshi Sifat Uz Zaman Department of Virology IEDCR Dhaka Bangladesh Department of Microbiology Popular Medical College Dhaka Bangladesh Department of Community Medicine Mugda Medical College Dhaka Bangladesh Department of Virology BSMMU Dhaka Bangladesh Infection Prevention and Control Division Medlife Healthcare Limited Dhaka Bangladesh

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.

关键词： CHIKV chikungynea RT-LAMP Infectious Diseases Viral Infection CHIKV-RNA

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