Comparative evaluation of two automated systems for nucleic acid extraction of BK virus: NucliSens easyMAG versus BioRobot MDx.
The objective of this study was to compare the performance of two automated nucleic acid extraction systems. Specifically, the NucliSens easyMAG system (bioMerieux, Marcy l'Etoile, France), which incorporates magnetic bead technology, was compared with the BioRobot MDx system (Qiagen GmbH, Hilden, Germany), which uses a silica membrane-based method of nucleic acid extraction. Nucleic acids from the BK virus (BKV DNA) were extracted from 98 plasma and 57 urine specimens using the Real-Q BKV quantitation kit (Biosewoom, Seoul, Korea). Failed PCR was defined as negative BKV DNA results having more than 36 threshold cycles of the internal control by the manufacturer's instruction. The PCR failure rate of nucleic acids isolated from plasma samples using the MDx system was similar to that of plasma samples processed using the easyMAG system (2.0% and 3.1%, respectively). The PCR failure rate of nucleic acids isolated from urine samples using the MDx system was higher than that of urine samples processed using the easyMAG system (33.3% and 12.5%, respectively). These data suggest that the PCR inhibitors present in urine specimens are removed more efficiently by the easyMAG system. Among amplified specimens, the discordant results obtained from the two systems revealed that the BKV DNA load ranged from 2.3 log10 copies/mL to 4.6 log10 copies/mL. Of the 25 urine specimens that yielded BKV DNA by both extraction systems, 15 specimens (60.0%) yielded higher BKV DNA loads by the easyMAG system, indicating that the easyMAG system extracted nucleic acid more efficiently than did the MDx system. In conclusion, the easyMAG method outperformed the MDx method when used to extract BKV DNA from urine samples. Magnetic bead-based extraction methods such as the easyMAG system are therefore preferable for the quantitation of viral DNA in urine.
Kim
Sollip
S
Department of Laboratory Medicine, University of Ulsan College of Medicine and Asan Medical Center, Asanbyeongwon-gil 86, Songpa-gu, Seoul 138-736, South Korea.
Park
Sook-Ja
SJ
Namgoong
Seung
S
Sung
Heungsup
H
Kim
Mi-Na
MN
D000328:Adult; D001331:Automation; D001739:BK Virus / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000097:blood / Q000302:isolation & purification* / Q000652:urine; D005260:Female; D006801:Humans; D016030:Kidney Transplantation / Q000009:adverse effects; D008280:Magnetics; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction / Q000379:methods*; D027601:Polyomavirus Infections / Q000175:diagnosis / Q000821:virology; D011933:Reagent Kits, Diagnostic; D033581:Stem Cell Transplantation / Q000009:adverse effects; D014412:Tumor Virus Infections / Q000175:diagnosis / Q000821:virology
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic