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Parallel evaluation of broad virus detection methods.

Jens Modrof1, Andreas Berting1, Thomas R Kreil2

  • 1Global Pathogen Safety, Baxter BioScience, Vienna, Austria.

PDA Journal of Pharmaceutical Science and Technology
|December 6, 2014
PubMed
Summary
This summary is machine-generated.

Broad virus detection methods show promise for biological product safety testing, accurately identifying most samples. However, further improvements are needed to minimize false positives and negatives before routine implementation.

Keywords:
Adventitious agent testingBlinded studyBroad virus detectionMassively parallel sequencingMicroarray

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Area of Science:

  • Virology
  • Biotechnology
  • Analytical Chemistry

Background:

  • Adventitious virus testing is crucial for biological product safety.
  • Emerging broad virus detection methods require evaluation for current procedures.

Purpose of the Study:

  • To assess the suitability of broad virus detection methods for adventitious agent testing.
  • To compare the performance of PCR-ESI/MS, microarray, and next-generation sequencing approaches.

Main Methods:

  • A blinded, comparative experimental study using spiked virus preparations.
  • Four different virus preparations were tested at two concentrations in two cell culture media.
  • Analysis performed using polymerase chain reaction-electrospray ionization mass spectrometry (PCR-ESI/MS), microarray, and two massively parallel sequencing methods.

Main Results:

  • All methods correctly identified the majority of samples (mean 83%), with a narrow range (72-95%).
  • Unexpected assignments (false positives) occurred in 20-45% of samples across methods.
  • False negatives were reported for 0-25% of samples; viruses detected at ~4-5 log10 qPCR copies/mL.

Conclusions:

  • Broad virus detection methods are suitable for detecting low virus concentrations.
  • Potential for false-positive and false-negative results necessitates further method refinement.
  • Methods require improvement before adoption into routine adventitious agent testing.