Comparison of the Isolation Methods of Viral Nucleic Acids

Year 2018, , 351 - 355, 30.09.2018

Nagehan Şakrucu Uğur Sıdal , Tamer Şanlıdağ

https://doi.org/10.18466/cbayarfbe.350005

Abstract

In vitro amplification of the nucleic acids (DNA
or RNA) is used in the detection of microbial agents and thus in the diagnosis
of infectious diseases, as well as in the diagnoses of oncological and genetic
disorders and forensic medicine. The aim of the present study was to compare
the isolation methods of the nucleic acids of hepatitis B and C viruses,
causative agents of the two significant infections worldwide. Conventional
isolation methods were compared with the commercial kits that have been used
commonly in recent years, in terms of reliability, cost-effectiveness,
contamination risk and duration of the testing time. Five standards for the
isolation of the viral nucleic acids of both HBV DNA (Fluorion HBV QNP 2.0) and
HCV RNA (Fluorion HCV QNP 2.1) were used. The isolations of the viral nucleic
acids of HBV and HCV were done with the conventional methods, phenol-chloroform
and guanidine thiocyanate, and the commercial kits Roboscreen and NucleoSpin. The
resultant viral nucleic acid load was determined with a spectrophotometer (WPA
UV 1101, Biotech Photometer), and their amplification was conducted with
Real-Time PCR. The results of the assessments revealed that the highest nucleic
acid concentration were obtained with the conventional methods, while they
exhibited significant drawbacks such as long duration of the testing time,
difficulty in application, and higher contamination risk.

Keywords

HBV, HCV, PCR, spectrophotometer, amplification.

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