Isolation methods of viral nucleic acids

Recent developments in molecular biology, especially nucleic acid amplification during the in vitro conditions, are frequently used for the diagnosis the diseases and for the forensic medicine. In amplification technique genetic materials are insulated from different organs, cambiums and fluids, are augmented by using enzymes. Amplification techniques are promptly, safe, sensitive, and specific but they aren?t affected by old materials, and timbleful factors or inactivity in body. Amplification methods are very beneficial to modify bacterial and viral agents that are insulate, generate, or identification very hardly. These techniques supply to economize on time and employees and minimize the employee fault and the laboratory infections altough costly, experienced, and learned employee and improved materials are needed (Arda,1995). Polymerase Chain Reaction, founded by RNA and DNA?s amplifications, is in the most used techiques. In this study, Hepatitis B and Hepatitis C, very important health problem over the world in recent years, amplifications are satisfied in PCR by making Hepatit B and Hepatit C viruses? insulations. There are two methods called conventional method and commercial kits, which are used in stage of virus insulation in the PCR technique. Our purpose is comparing reliability, loss of time, contamination, and cost between conventional method, having used previously, and commercial kits, whose usage become widespread. For the amplification of HBV DNA 5 standard (Fluorion HBV QNP 2.0) is used and for the amplification of HBV RNA (Fluorion HCV QNP 2.1 ) is used. Insulation for getting HBV RNA and HBV DNA is done by using the conventional method (Phenol-chloroform ve Guanidinium Thiocyanate method ) and kit method (Roboscreen, NucleoSpin ). After the insulation, measurements of total DNA and total RNA are done with the spectrophotometer. Maximum value of HBV DNA is confirmed between 90?g/ml -107,5 ?g/ml by Phenol-chloroform method. Maximum value of HBV RNA is determined 86?g/ml -108?g/ml by Guanidinium Thiocyanate method. Afterwards, satisfied amplification of DNA and RNA sources with PCR. According to spectrophotometer and PCR results, a comparison between conventional and commercial kit methods and modified advantages and disadvantages of these methods. Peak values are produced by conventional method but as to time insulation by kit is more easy and quick. Conventional method is more risky than kit method in contamination. Key words : HBV, HCV, Polymerase Chain Reaction, Spectrophotometer, Amplification.