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Zika Virus In America

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The Zika virus is rapidly becoming a prominent problem throughout the Americas, Africa, and Asia. The Zika virus is an arbovirus that inhabits Aedes species of mosquito populations and can infect humans through a bite. The Zika virus is also spread through sexual contact which is also causing fears in countries which Zika is rapidly spreading. The first case of Zika virus was discovered in Uganda in 1947 and has traveled across countries ever since. Zika is a single stranded RNA virus that has around 11000 base pairs. Upon infection, some common symptoms are fever, headache, rash, and diarrhea. These symptoms usually persist up to seven days and are usually mild in nature. However, some people may experience no symptoms at all which could further …show more content…

The Zika virus stocks that were grown were used to test the specificity and sensitivity of the rRT-PCR assay. Along with the Zika virus, other flaviviruses and arboviruses were used to test the specificity of this PCR reaction. The viral stocks were then titrated, stained and dried at room temperature. The plaques were counted once dried. After sequencing the Zika strains from Africa, the primers and LNA probe sequences were designed. The taqman probe contained a reporter dye, FAM, at the 5’ end with a quencher dye, TAMRA, at the 3’ end of the probe. The RNA quantification was done using the fluorescent dye RiboGreen was is specific to single-stranded RNA. A RNA standard that was used for the rRT-PCR was generated to identify the limit of the assay. For the extraction of the viral RNA, the RNA was extracted from the stock and mixed with buffer and ethanol and transferred to a column containing silica. The RNA was then amplified using real-time RT-PCR using a 25 ul reaction mixture including enzyme mixture, buffer, primers, probe, water and the RNA sample. The quantification was conducted in a 96-well plate used as a fluorimeter, to measure fluorescence. One positive and one negative control were used for each amplification. The positive control was stock nucleic acid and the negative control was water. The first cycle of PCR consisted of 10 minutes at 50°C and 15 minutes at 95°C. After the first cycle, 40 consequent cycles of 15 seconds at 95°C and then 1 minutes at 60°C. All 37 strains of the Zika virus were detected by the assay. The regression coefficient (R²=.9987) of the Ct value vs. Log graph indicated that the assay was highly reproducible. The detection limit for the rRT-PCR was shown to be .05 pfu per reaction using the same methods and materials. The Ct values ranged from 25.60 to 36.24 and were extremely correlated (R²=.9984). These

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