Mink coronavirus (MCoV) infection poses a serious threat to animal health, resulting in substantial economic losses in mink farming and impeding the sustainable development of the fur industry. Thus, developing both sensitive and specific molecular diagnostics is imperative for effective MCoV surveillance and disease control. In the present study, specific oligonucleotide primers were designed to target the 1b and N genes of MCoV, and two SYBR Green-based RT-qPCR assays - designated as MCoV-1b and MCoV-N, respectively - were established. The assays exhibited extremely high sensitivity, with lower limits of detection of 1.79×10¹ copies/μL for MCoV-1b and 1.29×10¹ copies/μL for MCoV-N. No cross-reactivity was observed with other common viruses and melting curve analyses revealed a single sharp peak for each assay, confirming specific amplification of MCoV targets. Reproducibility across three independent experimental runs exhibited low standard deviations and coefficients of variation of cycle threshold ( Ct ) values across broad dynamic ranges, indicating stable and consistent performance (CV < 5%). When applied to faecal clinical samples (n=236), the assays yielded overall positivity rates of 77.97% for the 1b target and 77.54% for the N targets, which are consistent with that from high-throughput sequencing, validating their utility. Collectively, the two RT-qPCR assays developed in this study offer a reliable technical platform for rapid screening, accurate quantification, and epidemiological monitoring of MCoV. The reported molecular diagnostics hold significant practical value in mitigating the risks of MCoV transmission, supporting the sustainable growth of the global mink farming industry and diminishing potential global public health threats. • Two SYBR Green RT-qPCR assays targeting 1b and N genes of mink coronavirus (MCoV) were developed and rigorously validated for single-copy sensitivity and absolute specificity. • No cross-reactivity with four common mink viruses - Aleutian disease virus, calicivirus, sapovirus, and norovirus - and displayed excellent reproducibility (CV < 5%). • Detected MCoV in 77.97% (1b) and 77.54% (N) of 236 faecal samples, fully concordant with high-throughput sequencing data. • The integrated platform enables rapid, cost-efficient screening and outbreak response, thereby facilitating standardized, evidence-based control of this economically consequential pathogen.
Niu et al. (2026) studied this question.