Rapid Detection of the H275Y Oseltamivir Resistance Mutation in Influenza A/H1N1 2009 by Single Base Pair RT-PCR and High-Resolution Melting
2011

Detecting Oseltamivir Resistance in Influenza A/H1N1 2009

Sample size: 69 publication Evidence: moderate

Author Information

Author(s): Tong Steven Y. C., Dakh Farshid, Hurt Aeron C., Deng Yi-Mo, Freeman Kevin, Fagan Peter K., Barr Ian G., Giffard Philip M.

Primary Institution: Menzies School of Health Research, Charles Darwin University

Hypothesis

We aimed to design a real-time reverse-transcriptase-PCR (rRT-PCR), high-resolution melting (HRM) assay to detect the H275Y mutation that confers oseltamivir resistance in influenza A/H1N1 2009 viruses.

Conclusion

The HRM assay can rapidly and cost-effectively screen samples for oseltamivir resistance in influenza A/H1N1 2009, although it is less sensitive than TaqMan assays.

Supporting Evidence

  • The HRM assay was validated on 69 influenza positive clinical samples.
  • The assay was able to differentiate between wildtype and mutant strains.
  • The study found that 34% of clinical samples would not be amenable to the HRM assay due to low viral concentrations.

Takeaway

Scientists created a test to quickly find out if a flu virus is resistant to a medicine called oseltamivir, which helps treat the flu.

Methodology

The study used a real-time reverse-transcriptase-PCR (rRT-PCR) followed by high-resolution melting (HRM) analysis to detect the H275Y mutation.

Limitations

The assay cannot reliably interrogate clinical samples with low amounts of viral RNA template.

Participant Demographics

Patients with influenza-like illness from Royal Darwin Hospital.

Statistical Information

P-Value

p<0.0001

Confidence Interval

95%

Statistical Significance

p<0.0001

Digital Object Identifier (DOI)

10.1371/journal.pone.0021446

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