Journal of agricultural and food chemistryOct 9, 2025

Rapid detection of PRRSV-1 using a combined one-step genetic test with easy-to-read results

CRISPR Gene Editing Weekly Brief ↗PubMed ↗DOI ↗

Updated Jun 28, 2026

Abstract

Essence

A one-tube RPA-CRISPR/Cas13d platform enabled rapid PRRSV-1 detection with lateral flow or fluorescence readouts in about 35 minutes.

Evidence

This diagnostic platform study developed an extraction-free, lyophilized CRISPR/Cas13d assay and, in clinical evaluation against RT-qPCR, reported 100% sensitivity and 100% specificity with a sample-to-answer workflow within 35 minutes.

Caveat

The abstract does not report the clinical sample size or evaluation breadth, which limits how firmly the reported perfect accuracy can be generalized.

Simplified

Rapid and scalable diagnostic technologies are essential for controlling infectious diseases. We present STEP (Single-Tube Extraction-free Platform for CRISPR/Cas13d detection), a streamlined, equipment-minimal CRISPR-based platform enabling rapid, sensitive, and accurate viral RNA detection. STEP integrates isothermal RPA amplification with CRISPR-based nucleic acid cleavage, providing multimodal readouts including lateral flow strips, in-tube fluorescence, and fluorescence quantification. Lab-free extraction reagents and lyophilized formulations enhance user friendliness of STEP and stability for point-of-care testing (POCT), reduce cost, and eliminate cold-chain requirements. Optimization of baseline time and the ability to operate at both ambient and body temperatures minimize temporal and equipment constraints. Clinical evaluation showed 100% sensitivity and specificity versus RT-qPCR, delivering a sample-to-answer workflow within 35 min. STEP provides a robust platform for decentralized infectious disease diagnostics and rapid public health response, combining speed, user friendliness, and minimal instrumentation requirements.