Comparison of manual and automated (MagNa Pure) nucleic acid isolation methods in molecular diagnosis of HIV infections HIV enfeksiyonunun moleküler tanisinda manuel ve otomatize (MagNa Pure) nükleik asit izolasyon yöntemlerinin karşilaştirilmasi


ALP A., Us D., Hasļik G.

Mikrobiyoloji Bulteni, vol.38, no.1-2, pp.77-83, 2004 (Scopus) identifier identifier

  • Publication Type: Article / Article
  • Volume: 38 Issue: 1-2
  • Publication Date: 2004
  • Journal Name: Mikrobiyoloji Bulteni
  • Journal Indexes: Scopus, TR DİZİN (ULAKBİM)
  • Page Numbers: pp.77-83
  • Keywords: Cobas Amplicor, Human immunodeficiency virus (HIV), Magna Pure, Nucleic acid isolation, Polymerase chain reaction
  • Hacettepe University Affiliated: Yes

Abstract

Rapid quantitative molecular methods are very important for the diagnosis of human immunodeficiency virus (HIV) infections, assessment of prognosis and follow up. The purpose of this study was to compare and evaluate the performances of conventional manual extraction method and automated MagNA Pure system, for the nucleic acid isolation step which is the first and most important step in molecular diagnosis of HIV infections. Plasma samples of 35 patients in which anti-HIV antibodies were found as positive by microparticule enzyme immunoassay and confirmed by immunoblotting method, were included in the study. The nucleic acids obtained simultaneously by manual isolation kit (Cobas Amplicor, HIV-1 Monitor Test, version 1.5, Roche Diagnostics) and automated system (MagNA Pure LC Total Nucleic Acid Isolation Kit, Roche Diagnostics), were amplified and detected in Cobas Amplicor (Roche Diagnostics) instrument. Twenty three of 35 samples (65.7%) were found to be positive, and 9 (25.7%) were negative by both of the methods. The agreement between the methods were detected as 91.4%, for qualitative results. Viral RNA copies detected by manual and MagNA Pure isolation methods were found between 76.0-7.590.000 (mean: 487.143) and 113.0-20.300.0000 (mean: 2.174.097) copies/ml, respectively. When both of the overall and individual results were evaluated, the number of RNA copies obtained with automatized system, were found higher than the manual method (p<0.05). Three samples which had low numbers of nucleic acids (113, 773, 857, respectively) with MagNA Pure, yielded negative results with manual method. In conclusion, the automatized MagNA Pure system was found to be a reliable, rapid and practical method for the isolation of HIV-RNA.