Background: EHV-1 and EHV-4 are common in horses globally, causing respiratory issues. EHV-1 can lead to severe signs, neurological disorders, and abortions. This affects sport horse performance and breeding, causing significant economic loss. Hence, a sensitive and specific diagnostic tool is crucial during outbreaks to determine which herpesvirus is involved. Objectives: To develop and evaluate a freeze-dried triplex qPCR kit for EHV-1 and EHV-4 diagnosis, ID Gene Lyo™ Equine Herpes virus 1 and 4 Triplex,1 containing an exogenous internal control. Study design: Assay evaluation. Methods: Limit of detection of the PCR (LDPCR) was determined with a synthetic nucleic acid of EHV-1 and EHV-4. The Method Detection Limit (MDL) was determined by using equine whole blood, respiratory swabs and foetal organ samples spiked with EHV-1 (ATCC® VR-700™2) and EHV-4 (ATCC® VR-2230™2) strains separately. Specificity was evaluated with strains from respiratory virus or bacteria from equine and other mammals. Results: The LDPCR (95%) was established around 3.12 copies/PCR for EHV-1 and EHV-4. The MDL obtained for EHV-1 was: 291.5 copies/mL for whole blood and respiratory swab, and 2915 copies/mL for foetal organ. The MDL obtained for EHV-4 was 473 600 copies/mL for whole blood, respiratory swab, and foetal organ. The new qPCR kit has no cross-reactions with other pathogens except EHV-8, which is closely related to EHV-1.[1] Main limitations: Not applicable. Conclusion: The new freeze-dried triplex qPCR kit for EHV-1 and EHV-4 kit allows for rapid detection of both viruses simultaneously, with high reliability. It can be used for different forms of equine herpesvirus infections involving many different sample types thanks to the exogenous internal control included. It is freeze-dried, allowing for easy shipment at ambient temperature worldwide, reducing shipping costs and the environmental footprint. Key manufacturers: 1 ID Gene Lyo™, https://www.innovativediagnostics.com; 2 https://genomes.atcc.org/ Ethical animal research: Not applicable. Competing interests: OM, AC, KK and PP are employees of IDvet. Informed consent: Not applicable. Funding: IDvet. Reference: [1] Garvey M, Suárez NM, Kerr K, Hector R, Moloney-Quinn L, Arkins S, Davison AJ, Cullinane A. Equid herpesvirus 8: Complete genome sequence and association with abortion in mares. PLoS One 2018;13(2):e0192301. 10.1371/journal.pone.0192301