AAVLD/USAHA 2021 Annual Meeting

Meeting today’s diagnostic needs—innovating for tomorrow’s challenges

At Thermo Fisher Scientific, we focus on customer needs and animal health. Our passion drives us to innovate and improve conditions for farm animals and the people who care for them.

Join us at the AAVLD/USAHA annual conference (Booth #100) to meet product experts and learn about new solutions your lab can utilize to meet the ever-changing demands in the field of veterinary diagnostics.

Poster

Improved Detection of T. foetus by RT-qPCR Eliminates the Need of Culture Medium

Denisse Meza, Megan Schroeder, Ivan Leyva Baca, and Rick Conrad

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Abstract: Tritrichomonas foetus (T. foetus) is a protozoan that is the causative agent of bovine trichomoniasis, a sexually transmitted disease found worldwide in bulls and cows. The previous DNA-based Trich detection kit, a quantitative PCR (qPCR) assay, has been considered the gold standard for PCR-based detection of T. foetus in preputial samples (smegma) collected in culture medium. However, in 2018, the Texas Veterinary Medical Diagnostic Laboratory (TVMDL) published primers and probe sequences that target the 5.8S ribosomal RNA rather than the gene, which generates earlier Ct values due to higher concentration of target template when compared to the DNA-targeting qPCR test. Since combining the reverse-transcription qPCR (RT-qPCR) primers and probe design with a one-step RT master mix makes the reaction more sensitive, it eliminates the need to collect and incubate smegma in the InPouch™ commercial culture medium and offers a simple PBS sample collection instead. Additionally, due to the higher target template concentration and the reduction in inhibition, it provides the capability to pool several smegma samples for nucleic acid extraction and RTqPCR.

For Research Use Only. Not for use in diagnostic procedures.

Presentation

Development of Updated Porcine Reproductive and Respiratory Syndrome Virus Products

Speaker: Mazen Ismail, R&D Scientist
Time: October 24, 1:45–2:00 pm, Room: Redrock 6

Abstract: Porcine reproductive and respiratory syndrome (PRRS) is a viral disease that leads to lower reproductive performance in breeding animals and respiratory disease in pigs. Due to the highly mutating nature of PRRSV and newly circulating strains, a project was launched to update the currently on market PRRSV products with a new molecular design to provide improved detection toward the new variants. The new reagents and controls provide a simpler molecular design which is based on VetMAX PRRSV EU & NA 2.0 assay available in Europe. A new molecular design was constructed after bioinformatically analyzing nearly 1600 sequences from public and private databases as well as testing nearly 400 positive field samples in vitro. Testing results displayed the ability of the new reagents to generate more accurate calls for PRRSV Type II compared to original PRRSV reagents. The new products have been externally tested by multiple collaborator labs and have generated concordant calls with the original products in 94.6% of samples and generated more accurate calls for 5.15% of total tested samples. The new reagents and controls significantly simplify the testing workflow by using a 2 step master mix compared to the 4 step workflow used by on market PRRSV reagents and controls. The updated products run in FAST mode and drop the run time down from 1 hour and 45 minutes to 49 minutes per run. These products are for research use only and are not intended to be used in diagnostic procedures.

Poster

Improved Detection of T. foetus by RT-qPCR Eliminates the Need of Culture Medium

Denisse Meza, Megan Schroeder, Ivan Leyva Baca, and Rick Conrad

Abstract: Tritrichomonas foetus (T. foetus) is a protozoan that is the causative agent of bovine trichomoniasis, a sexually transmitted disease found worldwide in bulls and cows. The previous DNA-based Trich detection kit, a quantitative PCR (qPCR) assay, has been considered the gold standard for PCR-based detection of T. foetus in preputial samples (smegma) collected in culture medium. However, in 2018, the Texas Veterinary Medical Diagnostic Laboratory (TVMDL) published primers and probe sequences that target the 5.8S ribosomal RNA rather than the gene, which generates earlier Ct values due to higher concentration of target template when compared to the DNA-targeting qPCR test. Since combining the reverse-transcription qPCR (RT-qPCR) primers and probe design with a one-step RT master mix makes the reaction more sensitive, it eliminates the need to collect and incubate smegma in the InPouch™ commercial culture medium and offers a simple PBS sample collection instead. Additionally, due to the higher target template concentration and the reduction in inhibition, it provides the capability to pool several smegma samples for nucleic acid extraction and RTqPCR.