The Design and Synthesis of a Heterologous RT-PCR Competitor for Quantitation of Immune System Messengers in Peripheral Blood Mononuclear Cells: Applications for the Study of Hepatitis C
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Hepatitis C virus (HCV) is a bloodborne pathogen that infects 4 million people in the United States. Most cases of HCV result in a chronic infection and liver damage. A small group of people exist who were exposed to HCV, but cleared the virus from their bodies without receiving treatment. Factors involving the innate component of the immune system in this "self cure" group may differ from those of chronic HCV patients. The Interferon system is a part of the nonspecific immune system which produces an anitviral response in healthy cells, enabling them to better resist infection. This projects aim is to facilitate the study of this aspect of the immune system in chronic HCV patients and the self cure group. Our approach is to compare the levels of interferon α 1 (IFNα 1), interferon α2 (IFNα2) and interferon 𝛽 (lFN𝛽), as well as 2'5' oligoadenylate sythetase (2'5' OAS) mRNA in peripheral blood mononuclear cells (PBMC) when exposed to virus from both groups of patients. In order to make a qualitative comparison, a reverse transcription polymerase chain reaction heterologous competitor was designed and synthesized by overlapping extension PCR (OE-PCR). This DNA competitor molecule will allow for precise quantitation of mRNA levels of the IFN's and 2'5' OAS and thus the comparison of the Interferon system and immune system of chronic hepatitis C patients and self-cure patients. Understanding this difference can lead to better, more effective Interferon therapies for patients infected with Hepatitis C.