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New Veterinary in vitro Diagnosis (IVD) using the Doctor Vida® Platform
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Testes “point-of-care” (POCT) são caracterizados como o diagnóstico junto dos espécimes. Este tipo de diagnóstico tem adquirido cada vez mais atenção do ramo veterinário devido ao aumento de zoonoses, tal como a COVID-19. POCT baseados em tecnologias de amplificação de ácidos nucleicos isotérmicas têm ganho popularidade por consumirem menos recursos que a PCR. O Doctor Vida® Pocket (DV Pocket) é um dispositivo POCT desenvolvido pela STAB VIDA que faz uso de LAMP em tempo real para o diagnóstico de COVID-19. Neste estudo desenvolveram-se 2 novos POCT, utilizando a plataforma DV Pocket, para diagnóstico da Disenteria Suína (DS) e Sexagem de Aves. DS é uma doença muco-hemorrágica causada por B. hyodysenteriae, afetando suiniculturas. Devido à rápida infeção por fezes e resistência a antibióticos pode causar grandes perdas financeiras, daí a importância de um POCT para o seu diagnóstico. Um teste baseado em LAMP, usando o gene nox de B. hyodysenteriae como marcador e E. coli como controlo, bem como 2 kits de colheita de fezes, foram desenvolvidos. Este apresentou uma sensibilidade de 78.9% e especificidade de 45.5% para o kit 1, 68.4% e 72.7% para o kit 2, sendo detetado dentro de 40 minutos, com apenas 2 minutos de hands-on. Sexagem de aves é essencial para estudos de conservação e evolução. A STAB VIDA possui serviços de sexagem de aves, desta forma este projeto visa facilitar o processo para os clientes. Testou-se em aves da família Psittacidae, Columba livia e Psittacula krameri. Neste último obtiveram-se os melhores resultados, com uma sensibilidade de 80%, dentro de 90 minutos de teste, com 30 minutos para incubação da amostra. No entanto, otimizações são ainda necessárias para ambos os testes de modo a que possam prosseguir para validação clínica, e assegurando uma maior sensibilidade e especificidade.
Point-of-Care Testing (POCT) is characterized as the diagnosis near the specimen. POCT devices in veterinary practice are getting more attention due to the increasing spread of zoonotic diseases, such as COVID-19. POCT based on isothermal Nucleic Acid Amplification Technologies (NAATs) are gaining popularity for consuming fewer resources and time compared to PCR. Doctor Vida® Pocket (DV Pocket) is a POCT device developed by STAB VIDA that allows to perform LAMP in real-time for diagnosis of COVID-19. Herein we describe the development of two new in vitro veterinary POCT using the DV Pocket platform: Swine Dysentery (SD) and Bird sexing. SD is a mucohaemorrhagic disease caused by B. hyodysenteriae affecting swine. The spread of SD through faeces and antibiotic resistance often leads to financial loss, thus the importance of developing a POCT. A LAMP-based assay targeting the nox gene from B. hyodysenteriae and E. coli as the sample control was developed, as well as two different stool collection kits. Ultimately, the assay presented 78.9% sensitivity and 45.5% specificity using kit 1 and a 68.4% sensitivity and 72.7% specificity for kit 2 allowing to detect B. hyodysenteriae within 40-minutes with only 2 minutes hands-on. Bird sex determination is essential for conservation and evolutionism studies. STAB VIDA provides a Bird Sexing Service, so this project aimed to facilitate the process for its clients. It was attempted for birds from the Psittacidae family, Columba livia and finally Psittacula krameri. The latter had the best results, with a sensitivity of 80%, within 90 minutes and 30 minutes of sample incubation. Optimizations on both assays are still required before progressing to clinical validation, ensuring higher sensitivity and specificity.
Point-of-Care Testing (POCT) is characterized as the diagnosis near the specimen. POCT devices in veterinary practice are getting more attention due to the increasing spread of zoonotic diseases, such as COVID-19. POCT based on isothermal Nucleic Acid Amplification Technologies (NAATs) are gaining popularity for consuming fewer resources and time compared to PCR. Doctor Vida® Pocket (DV Pocket) is a POCT device developed by STAB VIDA that allows to perform LAMP in real-time for diagnosis of COVID-19. Herein we describe the development of two new in vitro veterinary POCT using the DV Pocket platform: Swine Dysentery (SD) and Bird sexing. SD is a mucohaemorrhagic disease caused by B. hyodysenteriae affecting swine. The spread of SD through faeces and antibiotic resistance often leads to financial loss, thus the importance of developing a POCT. A LAMP-based assay targeting the nox gene from B. hyodysenteriae and E. coli as the sample control was developed, as well as two different stool collection kits. Ultimately, the assay presented 78.9% sensitivity and 45.5% specificity using kit 1 and a 68.4% sensitivity and 72.7% specificity for kit 2 allowing to detect B. hyodysenteriae within 40-minutes with only 2 minutes hands-on. Bird sex determination is essential for conservation and evolutionism studies. STAB VIDA provides a Bird Sexing Service, so this project aimed to facilitate the process for its clients. It was attempted for birds from the Psittacidae family, Columba livia and finally Psittacula krameri. The latter had the best results, with a sensitivity of 80%, within 90 minutes and 30 minutes of sample incubation. Optimizations on both assays are still required before progressing to clinical validation, ensuring higher sensitivity and specificity.
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Palavras-chave
Point-of-Care Diagnosis Isothermal Amplification LAMP Swine Dysentery B. hyodysenteriae