A comparison of the efficiency of commercially available automated nucleic acid extraction platforms to detect a wide range of respiratory viruses from a bank of clinical samples  — ASN Events
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  3. A comparison of the efficiency of commercially available automated nucleic acid extraction platforms to detect a wide range of respiratory viruses from a bank of clinical samples 

A comparison of the efficiency of commercially available automated nucleic acid extraction platforms to detect a wide range of respiratory viruses from a bank of clinical samples  (#206)

Ineka Gow 1 , Shoo Peng Siah 1 , Angela Elmore 1 , Dylan Warby 1 , Damien J Stark 2 , Jock Harkness 2 , John R Melki 1 , Douglas S Millar 1
  1. Genetic Signatures, Sydney, NSW, Australia
  2. St Vincents Hospital, Sydney, NSW, Australia

Objective: Molecular diagnostic techniques (MDx) are becoming increasing prevalent in molecular microbiology laboratories and are especially useful for the detection of viral pathogens, which have traditionally been diagnosed by enzyme immunosorbent assays (EIA) or tissue culture. MDx are particularly useful due to the rapid turn around time and increased specificity achieved compared to conventional technologies. Automation of viral nucleic acids extraction for downstream detection techniques, usually PCR based, markedly reduces hands-on time for laboratory workers and improves workflow. We sought to compare a wide range of commercially available systems in their ability to detect a bank of respiratory samples.

Methods: Nucleic acids were extracted from 100 primary clinical samples using a range of commercially available platforms including the Qiasymphony (Qiagen), Kingfisher Flex (Thermo), EasyMag (Biomerieux), MagNApure 96 (Roche) and Nimbus (Hamilton). Samples were extracted according to the manufacturers recommendations and PCR carried out using the Genetic Signatures EasyScreen™ Viral respiratory detection kit. Amplification reactions were carried out on a Biorad (Hercules) CFX384 machine. Each batch of samples from each system were ran simultaneously on the same PCR plate to ensure no variation, thus amplification efficiency could be determined precisely for each extraction instrument.

Results: Clinical samples were ran on each platform in order to give a representation of samples that would be received by routine testing laboratories. Platforms differed markedly in terms of both positivity and inhibition rates. The full details of the results obtained from each system will be discussed.

Conclusion: A wide range of nucleic acid extraction platforms are available and differ in sample throughput, instrument and reagent costs, footprint and whether the platform is open or closed. The data demonstrated that variations were observed with each platform in terms of the sensitivity for the detection of commonly encountered respiratory viruses. Users should be aware that results will differ between extraction systems and that extraction of nucleic acids is just as important as down stream PCR performance.

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