Display Settings:

Format

Send to:

Choose Destination
See comment in PubMed Commons below
J Clin Microbiol. 1997 Feb;35(2):511-4.

Use of heat release and an internal RNA standard control in reverse transcription-PCR detection of Norwalk virus from stool samples.

Author information

  • 1Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030, USA.

Abstract

Norwalk virus (NV) and the Norwalk-like viruses are important human pathogens that cause epidemic acute viral gastroenteritis. Current techniques used to recover NV from clinical samples involve multistep viral extraction and elution procedures with subsequent viral detection by reverse transcription-PCR (RT-PCR). In this study, a simple method using heat to recover viral RNA from 45 stool samples was compared to a conventional viral RNA extraction technique, with subsequent analysis by RT-PCR. In addition, we used an internal RNA standard for the detection of inhibitors present in processed samples. Our results indicate that the use of heat to recover NV RNA from stool samples has a sensitivity for the detection of NV RNA that is similar to the more labor-intensive, time-consuming, conventional RNA extraction technique. The use of an RNA internal standard permits the detection of inhibitors present in processed samples, allowing the identification of false negatives. The standard we developed has the advantage of allowing differential detection between wild-type viral RNA and standard using internal oligoprobe hybridization.

PMID:
9003630
[PubMed - indexed for MEDLINE]
PMCID:
PMC229614
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for HighWire Icon for PubMed Central
    Loading ...
    Write to the Help Desk