Peer-reviewed veterinary case report
Development of a primer-probe energy transfer based real-time PCR for the detection of Swine influenza virus.
- Journal:
- Journal of virological methods
- Year:
- 2013
- Authors:
- Kowalczyk, Andrzej et al.
- Affiliation:
- The National Veterinary Research Institute
Plain-English summary
Researchers have developed a new test to detect the swine influenza virus (SIV), which can affect both pigs and humans. This test uses a special technique called real-time reverse transcription-polymerase chain reaction (RT-PCR) that can identify the virus's genetic material in samples taken from infected animals. It is very sensitive and can detect even small amounts of the virus, making it useful for spotting outbreaks early, as it can find the virus just two days after infection. This new method could be a better option for diagnosing SIV compared to current tests. Overall, the new test appears to work well for detecting the virus in clinical samples.
Abstract
Swine influenza virus (SIV) causes a contagious and requiring official notification disease of pigs and humans. In this study, a real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on primer-probe energy transfer (PriProET) for the detection of SIV RNA was developed. The assay uses matrix gene-specific primers and an Oregon Green-labeled fluorescent probe and was employed for the detection of SIV in clinical samples to identify outbreaks and to monitor the prevalence of disease. The PriProET technology was used to obtain a probe melting profile for confirmation of the specific product amplification. The assay is specific for influenza virus with a sensitivity of detection limit of approximately 10 copies of RNA by PCR. Based on serial dilutions of SIV, the detection limit of the assay was approximately 0.003 TCID(50)/ml for H1N1 A/Swine/Poland/KPR9/2004 virus. The PriProET RT-PCR was suitable for the detection of SIV RNA isolated directly from clinical samples. The assay detected SIV RNA in pre-clinical swab samples as early as 2 days post-infection (dpi). The PriProET RT-PCR assay is an alternative to the existing diagnostic assays and could have enhanced applicability for clinical diagnosis.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/22944078/