Validation of an influenza virus A 5’Taq nuclease assay for the detection of equine influenza virus A RNA in nasal swab samples.

This research paper discusses the process of validating an influenza virus type A 5’Taq nuclease assay for detecting equine influenza virus A RNA in nasal swabs. The research concludes that this method of detection shows 98% agreement with a previous method and is thus a valid tool for detecting equine influenza virus.

Methodology

• The researchers used an in-house validation of a previously reported influenza virus type A 5’Taq nuclease assay. This is a test used to detect the presence of the equine influenza virus A RNA in nasal swab samples.
• They compared the efficiency of this test with a gel-based reverse transcription nested polymerase chain reaction (PCR), which was previously reported by the Irish Equine Centre. They chose this comparison as it targets a different region of the viral genome. Therefore, it is not just a repetition of the same test in a different format.
• The comparison with nested PCRs is useful as they are generally considered to have similar sensitivity to real-time PCRs and are therefore ideal for evaluation comparisons.

Results

• The research showed that the sensitivity of the nested PCR was comparable to that of the 5’Taq nuclease test.
• Known positive samples and known negative samples tested with both methods resulted in a 100% correlation.
• In parallel testing of 276 nasal swab samples, there was an agreement rate of 98% between the two testing methods.

Conclusions

• The researchers were able to confirm the specificity of nested amplicons through nucleotide sequencing.
• The sequencing showed over 99.5% identity with the same region of previously published equine influenza virus A sequences.
• Based on their findings, the researchers concluded that the real-time PCR is adequately validated as a reliable tool for detecting equine influenza A virus in equine nasal swabs.