CHIKV Case Study

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As stated in the beginning of this paper, CHIKV has similarities in signs and symptoms with Dengue Virus. Scientists were trying to formulate a more sensitive and specific approach to appropriately diagnose CHIKV. Currently, scientists are developing ways to better differentiate Zika, Dengue and CHIKV as they bear similarities in signs and symptoms. Despite having different genus species, they are similar in envelope protein folding and membrane fusion mechanisms. Furthermore, scientists studied proteomic sequencing provided by the National Center for Biotechnology Information (NCBI) protein database and researched on polymorphisms, potential protein fragments and possible antigenicity of peptides, and finally, calculated amino acid …show more content…

Extracted dengue viral RNA was converted to cDNA using RNA amplification with a highly conserved primer pair, i.e.D1 and D2. Dengue virus sequences that were amplified and typed using second-round amplification with primer D1 and four serotype-specific primers are given in the table below followed by according to the protocol published elsewhere (Lanviotti et al., 1992). Similarly, according to the published protocol specific primers were used for the detection of CHIKV (CHIKF — 5′-ACCGGCGTCTACCCATTCATGT-3′, nt10237-10258 and CHIKR — 5′-GGGCGGGTAGTCCATGTTGTAGA-3′, nt10544-10566) targeting a 325 base region in the cDNA encoding E1 protein of CHIKV strain 653496 (Accession # AY424803) amplified the products using thermal cycler (Master cycler, Eppendorf, Germany) (Myers and Carey, 1967; Khan et al., 2002). The PCR products from the above reactions were analyzed on 2% agarose gel in the figure below. The sequences obtained were blasted against the NCBI database to ascertain the amplification of specific viral sequences. (A) Agarose gel analysis of DNA products from RT-PCR of CHIKV samples 1.100 bp DNA ladder, 2.CHIKV positive control, 3–5 patient sample, 6. Negative control.
(B) Agarose gel analysis of DNA products from RT-PCR of RNA samples isolated from dengue viruses.
(I) After amplification with consensus primers.
(II) After second round amplification with type specific primers TS1, TS2, TS3 and TS4.
1.100 bp DNA ladder, D1—dengue type 1, D2—dengue

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