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Related Experiment Videos

Applied technique for increasing calicivirus detection in shellfish extracts.

W Burkhardt1, G M Blackstone, D Skilling

  • 1US Food and Drug Administration, Gulf Coast Seafood Laboratory, Dauphin Island, AL 36528-0158, USA. Wburkhar@cfsan.fda.gov

Journal of Applied Microbiology
|July 31, 2002
PubMed
Summary

A novel tube-within-a-tube (TWT) device enhances the detection of RNA viruses in food and clinical samples. This method improves sensitivity for enteric RNA viruses, even with inhibitory substances present.

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Area of Science:

  • Molecular Biology
  • Virology
  • Food Safety

Background:

  • Detection of enteric RNA viruses is crucial for public health.
  • Inhibitory substances in clinical, environmental, and food samples can hinder accurate viral detection.
  • Existing reverse transcription-polymerase chain reaction (RT-PCR) methods face challenges with sample inhibitors.

Purpose of the Study:

  • To optimize the detection of enteric RNA viruses using RT-PCR in the presence of sample inhibitors.
  • To evaluate a novel tube-within-a-tube (TWT) device for enhanced viral detection.
  • To assess the TWT device's performance in detecting calicivirus in various sample matrices.

Main Methods:

  • Adaptation of a TWT device for RT-PCR-nested PCR.
  • Detection of San Miguel sea lion virus (serotype 17) in water and oyster tissue extracts.

Related Experiment Videos

  • Comparison of the TWT device with conventional RT-PCR-nested PCR protocols.

    • Main Results:

    • The TWT device demonstrated a 10-fold decrease in the detection limit for calicivirus in shellfish tissue extracts compared to conventional methods.
    • The TWT device maintained an identical detection limit for viral nucleic acid in water samples.
    • Both methods estimated a particle-to-infectious particle ratio of approximately 40:1 for the tested calicivirus strain.

    Conclusions:

    • The TWT device shows promise for reducing the detection limit of caliciviruses and other RNA viruses in shellfish and potentially other food products.
    • The TWT approach is applicable for detecting low-level nucleic acid targets of bacteria or viruses in the presence of RT and/or PCR inhibitors in food and clinical specimens.