One-step narrow-thermal-cycling strand exchange amplification for sensitive detection of porcine reproductive and respiratory syndrome virus

May 22, 2025Archives of microbiology

Sensitive detection of pig reproductive and respiratory virus using a fast, single-step temperature cycling method

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Abstract

The detection limit of the new method for PRRSV is 6 copies µL of viral RNA.

A novel one-step method for detecting PRRSV was developed, showing high sensitivity and specificity.
The method can distinguish between currently circulating PRRSV genotypes without reacting to other porcine pathogens.
Detection time for the method is 35 minutes.
Clinical applicability was confirmed through testing of 5 inactivated blood samples and 62 clinical samples.
The method may also be useful for the rapid diagnosis of other RNA pathogens.

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Porcine reproductive and respiratory syndrome (PRRS), which is primarily characterized by respiratory and reproductive dysfunction, is an epidemic disease caused by porcine reproductive and respiratory syndrome virus (PRRSV) that has the potential to economically devastate the global swine industry. Rapid and accurate detection of PRRSV is critical for effective control of PRRS in swine. In this study, a novel one-step, highly sensitive and specific accelerated strand exchange amplification (ASEA) method for the detection of PRRSV was developed. The detection limit of the ASEA method was determined to be 6 copies µLof PRRSV RNA reference material or PRRSV in spiked swine blood. The ASEA method demonstrated the capacity to discern the currently circulating PRRSV genotypes without cross-reactivity with other porcine-derived pathogens that manifest similar clinical signs. The ASEA method exhibited a detection time of 35 min, and its clinical applicability was validated through the analysis of 5 inactivated blood samples and 62 clinical samples. The method's potential extends beyond the diagnosis of PRRSV, as it can also be applied to the rapid diagnosis of other RNA pathogens. This capacity is expected to make significant contributions to future epidemic prevention and surveillance efforts. -1

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One-step narrow-thermal-cycling strand exchange amplification for sensitive detection of porcine reproductive and respiratory syndrome virus

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