Comparison of manual and automated nucleic acid isolation methods for HBV-DNA and HCV-RNA assays

• Published in: Le infezioni in medicina Vol. 23; no. 3; pp. 247 - 252
• Main Authors: Yagmur, Gulhan, Altun, Hatice Uludag, Gökahmetoglu, Selma, Basok, Ela
• Format: Journal Article
• Language: English
• Published: Italy 01.09.2015
• Subjects: Automation - methods; DNA, Viral - isolation & purification; Hepacivirus - genetics; Hepatitis B - blood; Hepatitis B - diagnosis; Hepatitis B virus - genetics; Hepatitis C - blood; Hepatitis C - diagnosis; Humans; Nucleic Acid Amplification Techniques - methods; Reagent Kits, Diagnostic; Reproducibility of Results; RNA - isolation & purification; Sensitivity and Specificity; Specimen Handling - methods; Virology - methods
• ISSN: 1124-9390

In the diagnosis and monitoring of hepatitis B virus (HBV) and hepatitis C virus (HCV) infections, it is important to use methods that can provide rapid and reliable results. The present study aimed to compare the automated and manual extraction methods during the nucleic acid isolation phase for HBV-DNA and HCV-RNA assays. The study included 93 serum samples, 49 of which were for the HBV-DNA assay and 44 for the HCV-RNA assay. DNA and RNA isolation from the samples was performed manually with a "QIAmpMin Elute Kit" (Qiagen, Germany) and the automated isolation system, NucliSens easyMAG (BioMérieux, France). All the extraction products were amplified using the iCycler device (Bio-Rad, USA). With both methods, compliance was found in 21 (42.8%) samples in the HBV-DNA assay; nine (18.3%) samples had a higher amount of viral nucleic acid with the manual method, whereas 19 samples (38.7%) were found to have a higher amount of nucleic acid with the automated system. For the HCV-RNA assay, total compliance was found in 31 (70.4%) samples; 12 (27.2%) samples had a higher amount of viral nucleic acid with the manual method whereas one sample (2.2%) was found to have a higher amount of nucleic acid with the automated system. It was concluded that the NucliSens easyMAG automated isolation system can be used with confidence for nucleic acid extraction due to its higher sensitivity, providing results in a shorter time, and assured standardization.