A recombinase polymerase amplification assay can return a readable African swine fever virus result in under 10 minutes, at 37–42 °C, without a thermocycler — and a new review argues that capability is redefining where veterinary diagnostics happen.

The review, published in Frontiers in Veterinary Science on May 7, 2026, by Tooryan, Enyertornye, Choudhury and Velayudhan of the University of Georgia, synthesizes two decades of work on isothermal nucleic acid amplification across livestock and companion-animal medicine. Its economic framing is blunt: paratuberculosis costs affected US dairy herds roughly US$100 per cow on average and more than US$200 per cow in high-prevalence herds, with US$250 million in annual losses industry-wide. Mycobacterium tuberculosis complex and mastitis together cost the global cattle industry approximately US$35 billion, and African swine fever cost China an estimated US$111.2 billion in 2019 alone.

PCR remains the reference standard, but it depends on thermal cyclers, laboratory infrastructure, trained personnel and time-consuming protocols. Clinical specimens also carry inhibitors — heme, urea, heparin, immunoglobulin G, lactoferrin, polysaccharides — that degrade amplification. The authors note evidence that LAMP tolerates these inhibitors better than PCR. In one detailed evaluation of seven common inhibitors (bile salts, calcium chloride, hematin, humic acid, IgG, tannic acid, urea), the concentrations that affected LAMP were comparable to or higher than those impacting PCR, and end-point detection of amplicons remained largely unaffected.

How the platforms compare

The review organizes the field by mechanism. LAMP uses four to six primers recognizing six to eight target regions, runs at 60–65 °C with Bst polymerase, and delivers results in 30–45 minutes with colorimetric, fluorescent, turbidimetric or lateral-flow readouts. RPA runs at 37–42 °C and produces detectable product in 10–20 minutes — modest heat sources, even body heat, suffice. Rolling circle amplification (30–37 °C, 60–120 minutes) offers single-molecule sensitivity via Φ29 polymerase and padlock probes. NASBA targets RNA at about 41 °C using reverse transcriptase, RNase H and T7 polymerase. Multiple displacement amplification (~30 °C, 2–4 hours) amplifies whole genomes rather than specific targets. Helicase-dependent amplification (37–65 °C, 60–120 minutes) mimics natural replication. Newer entrants include cross-priming amplification and strand-exchange amplification, the latter running in 15–30 minutes with a single primer pair and one polymerase.

What the field data show

For food-animal practitioners, the performance evidence spans bacteria, viruses and parasites. LAMP assays targeting Salmonella genes (invA, bcfD, safA for Enteritidis, yeeA for Heidelberg) showed high specificity, though one LAMP-only approach achieved under 90% sensitivity in chicken feces; adding CRISPR-Cas12b detection significantly improved both sensitivity and specificity in that matrix. A LAMP–lateral flow immunoassay for mastitis-causing bacteria in bovine milk produced both false positives and false negatives — a caution that matrix and readout format drive reliability.

On the viral side, RT-LAMP for avian influenza virus matched real-time RT-PCR sensitivity in chicken field samples with no cross-reactivity against non-target avian or bovine pathogens. LAMP with lateral-flow readout for ASFV showed enhanced sensitivity relative to real-time PCR, ELISA and standalone LAMP. A graphene oxide–based accelerated SEA assay applied to ASFV in pig nasal swabs reduced nonspecific amplification by suppressing primer dimers. Saltatory RCA achieved complete specificity and higher analytical sensitivity than real-time PCR for ASFV in clinical samples.

For companion animals, RT-RPA assays provide simple, rapid canine distemper virus detection in laboratories and point-of-care facilities; the authors single out animal shelters, where rapid isolation and outbreak control are critical. LAMP has detected Toxoplasma gondii in cat blood, cat feces, dog blood and pet bird brain tissue, while RPA coupled with CRISPR/Cas12a lateral-flow assays detected T. gondii in blood from stray dogs and cats in Zhejiang province, China.

The CRISPR and microfluidics multiplier

Pairing isothermal amplification with CRISPR-Cas12 and Cas13 systems enables collateral-cleavage signal amplification and detection down to attomolar levels. CRISPR-enhanced LAMP has detected Brucella species directly from animal serum in under 40 minutes. Microfluidic cartridges integrating extraction, amplification and CRISPR detection run on ≤10 μL sample volumes in battery-powered devices; one centrifugal microfluidic disk identified six major swine viruses within an hour.

Contamination remains the field's structural weakness. Isothermal reactions generate large amplicon volumes, and opening tubes for lateral-flow readouts risks aerosolized carryover and false positives. Mitigations include closed-tube LED visualization, wax-sealed dyes, uracil-DNA glycosylase supplementation and CRISPR/Cas9-based contaminant destruction.

The public-health dimension runs through the review's One Health framing: portable, low-cost pathogen detection strengthens zoonotic surveillance, food safety and outbreak response, allowing veterinarians, inspectors and producers to trigger quarantine, treatment or product recalls before disease spreads.

None of the assays described carries FDA CVM, EMA or USDA licensure status in the review; the authors frame the technologies as research-stage or variably commercialized, with SEA in particular lacking commercial kits and clinical-scale validation.

The authors close on standardization: wider test standardization, real-world performance evaluation in complex matrices, and demonstrated cost-effectiveness in resource-poor settings remain the gates between today's published assays and routine deployment — but with inhibitor-resistant enzymes and expanding CRISPR-microfluidic multiplexing, they expect isothermal platforms to play a steadily larger role in veterinary diagnostics.