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A sensitive and robust method for measles RNA detection

N Chadwick1, I Bruce, M Davies

  • 1Inflammatory Bowel Disease Study Group, Royal Free Hospital School of Medicine, Hampstead, London, UK.

Journal of Virological Methods
|March 20, 1998
PubMed
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This study compared measles RNA amplification methods, finding hybrid capture with combined reverse transcription polymerase chain reaction (RT-PCR) most robust for tissue samples. Nucleic acid sequence-based amplification (NASBA) was most sensitive for water samples.

Area of Science:

  • Virology
  • Molecular Biology
  • Biotechnology

Background:

  • Measles virus RNA detection is crucial for diagnosis and research.
  • Optimizing RNA amplification methods enhances sensitivity and robustness.

Purpose of the Study:

  • To compare different measles RNA amplification and detection techniques.
  • To develop and select the most rapid, sensitive, and robust procedure for measles RNA detection.

Main Methods:

  • Evaluated hybrid capture for RNA isolation.
  • Compared three RNA amplification methods: nested RT-PCR, combined RT-PCR with rTth polymerase, and NASBA.
  • Developed an internal positive control for assay validation.

Main Results:

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  • Hybrid capture followed by combined RT-PCR with rTth polymerase demonstrated superior robustness and sensitivity in tissue homogenates.
  • This optimized protocol detected as few as 10^4 synthetic measles RNA transcripts.
  • NASBA exhibited the highest sensitivity for measles RNA detection specifically in water samples.

    • Conclusions:

    • Combined RT-PCR with hybrid capture offers a robust and sensitive method for measles RNA detection in biological tissues.
    • NASBA is a highly sensitive alternative for detecting measles RNA in aqueous solutions.
    • The developed internal control aids in assay reliability.