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Simple Summary: African swine fever virus is a pathogen capable of spreading among swine populations, which is usually fatal since no vaccines are currently available on the market. Outbreaks caused by this virus are the reason for massive economic losses on pig farms and are a matter of worry for the swine sector worldwide. Effective and reliable detection of the virus is relevant to prevent uncontrolled contagion, especially when no effective preventive measures can be applied. In order to confront the spread of African swine fever, several rapid real-time polymerase chain reaction detection assays were developed by different research groups. In the present study, we focused on the validation of the ASFV MONODOSE dtec-qPCR kit using reference genetic material provided by national and international reference laboratories, and results were compared with those obtained from using internationally accepted reference methods. The data obtained indicated that the kit can be used in veterinary samples. Technical innovations enable a user-friendly and rapid management of samples, reducing the probability of human errors happening and transport costs. African swine fever virus is considered an emerging virus that causes African swine fever, a disease characterised by high mortality and elevated transmission rates and that, as it is for most other viral diseases, cannot be treated with specific drugs. Effective and reliable detection of the virus is relevant to prevent uncontrolled contagion among boar populations and to reduce economic losses. Moreover, animal health laboratories are demanding standardisation, optimisation and quality assurance of the available diagnostic assays. In the present study, the ASFV MONODOSE dtec-qPCR kit was validated following the UNE-EN ISO/IEC 17025:2005 guidelines. Analytical validation terms include in silico and in vitro specificity, sensitivity, efficiency and reliability (repeatability/reproducibility). Diagnostic validation of the method was assessed through the analysis of a total of 181 porcine samples originating from six different matrix types doped with African swine fever virus DNA received from the European reference laboratory for African Swine Fever (INIA-CISA, Madrid, Spain): whole blood, blood serum, kidney, heart, liver and tonsil. Results agreed with those obtained from a reference detection method also based on real-time PCR, endorsed by WOAH, but the ASFV MONODOSE dtec-qPCR kit incorporates some technical innovations and improvements which may benefit end-users. This kit, available worldwide with full analytical and diagnostic validation, can recognise all known ASFV genotypes and brings additional benefits to the current qPCR technology. [ABSTRACT FROM AUTHOR]

Human mpox is caused by the Monkeypox virus, a microorganism closely related to the Variola virus, both belonging to the Orthopoxvirus genus. Mpox had been considered a rare disease until a global outbreak occurred in 2022. People infected with the virus present similar symptoms to patients suffering smallpox and other rash illnesses, hindering diagnosis. The WHO indicated that no commercial PCR or serology kits are currently widely available. In the present study, the MPXV MONODOSE dtec-qPCR kit was validated following guidelines of the UNE/EN ISO/IEC 17025:2005. The parameters evaluated for the acceptance of the assay were in silico and in vitro specificity, quantitative phase analysis, reliability, and sensitivity. The assay passed validation criteria and yielded an efficiency of 95.8%, high repeatability, reproducibility, and a Limit of Detection and Quantification of at least 10 copies. Results from the validation of the MPXV dtec-qPCR kit were satisfactory. The use of the MONODOSE format (dehydrated single PCR-tubes, ready to use) provided considerable advantages allowing the detection of the Monkeypox virus to be accurately achieved. This detection kit may be considered a reliable, fast, simple, and universally available option. [ABSTRACT FROM AUTHOR]

Summary: Candida auris is an emerging multidrug resistant pathogenic fungus that causes candidaemia with high mortality rates and exhibits the ability to persist within the hospital environment. Candida auris is phylogenetically closely related to Candida haemulonii, C. lusitaniae, C. pseudohaemulonii and C. duobushaemulonii and is frequently misidentified by commercial identification methods. In the present study, the GPS™MONODOSE dtec‐qPCR kit (dried single‐dose PCR tubes) for the detection of C. auris was validated following the guidelines of the UNE/EN ISO/IEC 17025:2005, the French Standard NF T90‐471:2010 and using fast‐cycling protocols. Validation terms included in vitro specificity (inclusivity/exclusivity), quantitative phase analysis (10‐106 standard DNA copies), reliability (repeatability/reproducibility) and sensitivity (detection/quantification limits). GPS™ dtec‐qPCR kits passed validation with strict acceptance criteria (n ≥ 10 repetitions). In silico specificity was proven by the designer (GPS™). Experimental inclusiveness was achieved in two independent laboratories by testing strain JCM15448T and 117 clinical C. auris isolates from Asia, the Middle‐East Africa, Latin America and Europe. Exclusiveness was evaluated with 25 strains of closely related Candida spp. Use of the MONODOSE format provided considerable advantages allowing the detection of C. auris to be accurately achieved in less than an hour. The GPS™MONODOSE dtec‐qPCR kit is ready to undergo clinical evaluation. [ABSTRACT FROM AUTHOR]