Molecular differentiation of dengue virus serotypes using RT-PCR and estimation of their effect on liver function

Dr. Abhinav Manish, P. Pandey, N. Sharma
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Abstract

Dengue is one of the common and leading cause of mortality and morbidity in tropical countries specially in India. Its hepatotoxic effects have been demonstrated by the researchers over a long time. Dengue virus is a RNA virus, and known to have four serotypes and recently a fifth variant is also demonstrated in India. The effect of the different serotypes is seldomely investigated by the researchers. So we try to estimate the effect on Liver due to infection via different serotypes of Dengue virus. After taking the ethical clearance, from the ethical committee, A hospital based prospective study was conducted from August 2017 to November 2018, on patients attending OPD and IPD, of Medicine Department at Shri Mahant Indiresh Hospital Dehradun. 60 Dengue Positive cases were selected. RNA gets extracted using Reverse Transcriptase PCR, Conventional PCR and Real Time PCR techniques and further visualized using Agarose gel electrophoresis techniques at the CMRL (Central Molecular Research Laboratory), and liver Enzymes SGPT, SGOT, ALP, GGT were analyzed at Central Clinical Laboratory of SMI Hospital using the vitros 5600 fully autoanlyzer of the Orthoclinical diagnostics. Results were analyzed using suitable statistical tools.
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利用RT-PCR技术鉴定登革病毒血清型及其对肝功能的影响
登革热是热带国家,特别是印度常见和主要的死亡和发病原因之一。它的肝毒性作用已经被研究人员证明了很长时间。登革热病毒是一种RNA病毒,已知有四种血清型,最近在印度也发现了第五种变体。研究人员很少调查不同血清型的影响。因此,我们试图估计不同血清型登革热病毒感染对肝脏的影响。在获得伦理委员会的伦理许可后,于2017年8月至2018年11月对德拉敦Shri Mahant Indiresh医院内科门诊和IPD的患者进行了一项基于医院的前瞻性研究,选择了60例登革热阳性病例。RNA提取采用逆转录酶PCR、常规PCR和Real Time PCR技术,并在中央分子研究实验室采用琼脂糖凝胶电泳技术进行进一步可视化,肝酶SGPT、SGOT、ALP、GGT在SMI医院中心临床实验室采用Orthoclinical diagnostics的5600全自动体外分析仪进行分析。采用合适的统计工具对结果进行分析。
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