Peer-reviewed veterinary case report
Reverse transcription recombinase polymerase amplification assay for the rapid detection of type 2 porcine reproductive and respiratory syndrome virus.
- Journal:
- Journal of virological methods
- Year:
- 2017
- Authors:
- Wang, Jian-Chang et al.
- Affiliation:
- Inspection and Quarantine Technical Center of Hebei Entry-Exit Inspection and Quarantine Bureau · China
Plain-English summary
Researchers have developed a new test to quickly detect a virus called type 2 porcine reproductive and respiratory syndrome virus (PRRSV), which is a major problem for pigs and the farming industry. This test, known as reverse transcription recombinase polymerase amplification (RT-RPA), can identify the virus in just 20 minutes at a specific temperature and is very sensitive, able to detect even small amounts of the virus. In tests with real pig samples, the RT-RPA method found the virus in 86.6% of cases, which is slightly better than another common test called real-time RT-PCR, which detected it in 83.3% of cases. This new method is simple and could be very useful for quickly diagnosing the virus in farms, especially in rural areas. Overall, the test appears to work well for detecting this important virus in pigs.
Abstract
Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in pigs, and has tremendous negative economic impact on the swine industry worldwide. PRRSV is classified into the two distinct genotypes: type 1 and type 2, and most of the described PRRSV isolates in China are type 2. Rapid and sensitive detection of PRRSV is of great importance for the disease control and regional eradication programs. Recombinase polymerase amplification (RPA) has emerged as a novel isothermal amplification technology for the molecular diagnosis of infectious diseases. In this study, a fluorescence reverse transcription RPA (RT-RPA) assay was developed to detect the type 2 PRRSV using primers and exo probe specific for the viral nucleocapsid gene. The reaction was performed at 40°C within 20min. The RT-RPA assay could detect both the classical (C-PRRSV) and highly pathogenic PRRSV (HP-PRRSV), but there was no cross-reaction to other pathogens. Using the in vitro transcribed PRRSV RNA as template, the analytical sensitivity of RT-RPA was 690 copies. The assay performance was evaluated by testing 60 field samples and compared to real-time RT-PCR. The detection rate of RT-RPA was 86.6% (52/60), while the detection rate of real-time RT-PCR was 83.3% (50/60). This simple, rapid and reliable method could be potentially applied for rapid detection of PRRSV in point-of-care and rural areas.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/28122203/