Development of dual DIVA real-time PCR to differentiate between virulent and vaccine strains of lumpy skin disease virus
Lu-Jen Ting
Abstract
Lumpy skin disease (LSD) is a notifiable disease of World Organisation for Animal Health (WOAH) due to its significant health and economic impact on cattle. LSD attenuated live vaccines have satisfactory protection and can effectively control the spread of the disease. However, in some cases the vaccine strain may cause mild or systemic skin reactions, which are indistinguishable from the infection of virulent field strain. In this study, it was found that the virulent virus carries 71 bases less than the vaccine strain in the ankyrin repeat protein gene and this feature allows to design specific primers and probes for each specific DIVA dual real-time polymerase chain reaction, to differentiate vaccine strain (by qPCR-Vac) from vaccine-like recombinant LSDVs (by qPCR-WV). It was verified that the detection ability of qPCR-Vac and qPCR-WV with the WOAH recommended capripoxvirus qPCR (qPCR-capri) was 0.92 and 0.97, respectively, indicating that the agreement were both high. The synthetic positive control oligonucleotide was detected by qPCR-Vac and qPCR-WV, both with sensitivity up to 10 copies. Therefore, the newly developed DIVA qPCRs are reliable to check clinical signs timely and to allow authorities to control the disease.
2023 International High-Containment Laboratory Practices and Techniques Seminar
Li-Hsuan Chen
Abstract
Kansas State University Biosecurity Research Institute (BRI) is a unique institute in high containment facilities for researching and training and integrates veterinary medicine, plant pathology, food safety, and molecular biology. The research activities of food-born, animal, plant, and arthropod-borne pathogens and zoonosis pathogens can be held in one building, allowing the BRI become a leader in agriculture and public health.
To promote safe work practices and techniques in high containment laboratory worldwide, Department of Agriculture Animal and Plant Health Inspection Service (APHIS) funded the international participants to attend ”International High-Containment Laboratory Practices and Techniques Seminar”. The content of the seminar included laboratory biosafety, risk assessment and mitigation strategies, personal protective equipment use, biosafety cabinet use, transportation of infectious materials, emergency procedures, etc. In the last lesson, participants are asked to organize a project on high-risk pathogens in BSL-3 by group presentation.