21034773
not annotated - annotated - LINNAEUS only
Comparison of commercial systems for extraction of nucleic acids from DNA/RNA respiratory pathogens.
This study compared six automated nucleic acid extraction systems and one manual kit for their ability to recover nucleic acids from human nasal wash specimens spiked with five respiratory pathogens, representing Gram-positive bacteria (Streptococcus pyogenes), Gram-negative bacteria (Legionella pneumophila), DNA viruses (adenovirus), segmented RNA viruses (human influenza virus A), and non-segmented RNA viruses (respiratory syncytial virus). The robots and kit evaluated represent major commercially available methods that are capable of simultaneous extraction of DNA and RNA from respiratory specimens, and included platforms based on magnetic-bead technology (KingFisher mL, Biorobot EZ1, easyMAG, KingFisher Flex, and MagNA Pure Compact) or glass fiber filter technology (Biorobot MDX and the manual kit Allprep). All methods yielded extracts free of cross-contamination and RT-PCR inhibition. All automated systems recovered L. pneumophila and adenovirus DNA equivalently. However, the MagNA Pure protocol demonstrated more than 4-fold higher DNA recovery from the S. pyogenes than other methods. The KingFisher mL and easyMAG protocols provided 1- to 3-log wider linearity and extracted 3- to 4-fold more RNA from the human influenza virus and respiratory syncytial virus. These findings suggest that systems differed in nucleic acid recovery, reproducibility, and linearity in a pathogen specific manner.
Ann file
T1 Species 236 241 human
N1 Reference T1 Taxonomy:9606
T2 Species 340 362 Streptococcus pyogenes
N2 Reference T2 Taxonomy:1314
T3 Species 389 411 Legionella pneumophila
N3 Reference T3 Taxonomy:446
T4 Species 463 486 human influenza virus A
N4 Reference T4 Taxonomy:11320
T5 Species 520 547 respiratory syncytial virus
N5 Reference T5 Taxonomy:12814
T6 Species 1038 1052 L. pneumophila
N6 Reference T6 Taxonomy:446
T7 Species 1178 1189 S. pyogenes
N7 Reference T7 Taxonomy:1314
T8 Species 1332 1353 human influenza virus
N8 Reference T8 Taxonomy:11320
T9 Species 1358 1385 respiratory syncytial virus
N9 Reference T9 Taxonomy:12814
T10 Species 427 437 adenovirus
N10 Reference T10 Taxonomy:10535
T11 Species 1057 1067 adenovirus
N11 Reference T11 Taxonomy:10535