Evalution of Dengue virus RNA extraction methods and the study of viral-induced apoptosis of HepG2 hepatocyte
Dengue fever is an arthropod-borne transmit disease, caused by dengue virus.The principal vectors are Aedes aegypti and Aedes albopictus. Induce Dengue fever (DF) and a more severe form of dengue hemorrhagic fever (DHF), dengue shock syndrome (DSS). Dengue virus is the most prevalent arbovirus in tropical and subtropical regions. There is no specific drug and vaccine available for treatment and prevention. Therefore, DF is an important disease among transmit diseases in humans. For the effective control and prevention of DF transmission, rapid quantitative molecular biological methods are very important for the diagnosis of dengue fever. At present, there are many methods to isolate the RNA of Dengue virus; however, the new developed magnetic method has not been used for the Dengue virus isolation yet. At first, we evaluated various methods for Dengue virus isolation. The result indicate that the best method of RNA extraction for dengue virus is the QIAamp® Viral RNA kit manual extraction. There are no apparent differences of the effect for Dengue virus RNA isolation between filting film and magnetic bead method. Furthermore, DHF caused by Dengue virus is a very serious disease and the pathologic mechanism of DHF has not been elucidated completely. Both clinical and experimental trials have confirmed that the liver cell is one of the target infected by Dengue virus. And, the mechanism of Dengue virus-induced liver cell apoptosis remains poorly understood.Furthermore, there are free radical and cytokines production in patient¡As serum in the acute phase of DF. Therefore, the role of antioxidant and p21 in the mechanisms should be elucidcited. Our preliminary data show that p21 mRNA expression increase in HepG2 after Dengue virus infection. NAC, GSH, and DPI all can attenuate Dengue-induced cell apoptosis. Howerer, the relationship between p21 expression and liver cell apoptosis should be further clarified in the near future.
Advisor:Jong-kang Lin; Huei-sheng Huang; Jiin-tsuey Cheng; Kung-chia Young
School:National Sun Yat-Sen University
School Location:China - Taiwan
Source Type:Master's Thesis
Keywords:p21 hepg2 cell antioxidants dengue
Date of Publication:09/04/2006