Comparison of the Isolation Methods of Viral Nucleic Acids
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Date
2018
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Abstract
In vitro amplification of the nucleic acids (DNAor RNA) is used in the detection of microbial agents and thus in the diagnosisof infectious diseases, as well as in the diagnoses of oncological and geneticdisorders and forensic medicine. The aim of the present study was to comparethe isolation methods of the nucleic acids of hepatitis B and C viruses,causative agents of the two significant infections worldwide. Conventionalisolation methods were compared with the commercial kits that have been usedcommonly in recent years, in terms of reliability, cost-effectiveness,contamination risk and duration of the testing time. Five standards for theisolation of the viral nucleic acids of both HBV DNA (Fluorion HBV QNP 2.0) andHCV RNA (Fluorion HCV QNP 2.1) were used. The isolations of the viral nucleicacids of HBV and HCV were done with the conventional methods, phenol-chloroformand guanidine thiocyanate, and the commercial kits Roboscreen and NucleoSpin. Theresultant viral nucleic acid load was determined with a spectrophotometer (WPAUV 1101, Biotech Photometer), and their amplification was conducted withReal-Time PCR. The results of the assessments revealed that the highest nucleicacid concentration were obtained with the conventional methods, while theyexhibited significant drawbacks such as long duration of the testing time,difficulty in application, and higher contamination risk.