Next-Gen Sequencing is an important diagnostic tool for veterinarians.

Next-Gen Sequencing is an important diagnostic tool for veterinarians.

DNA (also known as deoxyribonucleic acid) has long been nicknamed the “blueprint for life”, providing the genetic codes that make existence for all organisms possible [1]. From animals to bacteria to fungi, each organism’s genetic code is what makes them unique and can also provide an abundance of information for clinical uses. Specifically, the development of high throughput molecular technologies alongside bioinformatics analyses in the 21st century has radically changed the abilities of veterinarians to give their patients more informed diagnoses and treatments [2]. NGS is a “high-resolution tool that provides veterinary diagnostic laboratories with the ability to undertake swift and flexible responses to emerging infectious diseases and unexpected pathogen variants” [2].

If you suspect your pet is suffering from an infection, it is recommended that you make an appointment with your veterinarian to diagnose and provide a treatment plan tailored to your pet’s needs as soon as possible!

What can DNA tell veterinarians about your pet’s infection?

DNA is composed of two parallel strands made from a phosphate sugar backbone and is connected by chemical bonds between bases (adenine bonds to thymine, and cytosine bonds to guanine), which creates a double helix [1]. NGS has allowed for the characterization of complete microbial communities, even those without prior knowledge [3]! The analysis of metagenomics data can not only help identify and quantify the microbiome of interest, but can also be function based and identify coding gene diversity [2]. This means that NGS diagnostics can help screen for virulence associated, antibiotic resistance genes, and vitamin production-associated genes in the microbial communities [2]!

So, how does it work?

For NGS diagnostics, microbial, genomic DNA or RNA (converted to cDNA) is extracted from clinical samples, such as urine, feces, blood, or skin/ear/nose/throat swabs, then purified and tagged. The resulting DNA molecules are copied thousands of times and massively sequenced in parallel. Thus, millions of DNA sequence reads from all microbes in a sample are able to be examined at once. These DNA reads are then trimmed, host DNA is filtered out using special software, overlapping reads are assembled into long sequences and the genes are compared to genomic databases to assign the microbial species identity and associated pathogenicity and predict antimicrobial activity based on resistance genes. This process provides a complete picture of which microbes are present and allows the quantification of each microbial species at the time and sight of sample collection from the patient. NGS-based diagnostic tools are increasingly used in veterinary practices to further improve infection diagnostics, while also aiding good antibiotic stewardship by selecting the most useful antibiotics. These types of tests are offered by specialty labs, like the MiDOG labs pet microbiome test.

Next-Gen DNA Sequencing Technology in Veterinary Medicine

The image above depicts the MiDOG All-in-One Microbial Test workflow.

Why is Next-Gen Sequencing Better Than Cultures or PCR?

Historically, culture-based methods and PCR techniques have been used to assess potential emerging infections in veterinary medicine, but there are notable diagnostic shortcomings for both. Only one percent of all microbes can actually be cultured using standard culturing methods, which lack the sensitivity and ability to cultivate difficult to grow pathogens [4]. While PCR provides more rapid results, the diagnostic value of these tests is limited because they are only able to target a known set of pathogens.

The MiDOG All-in-One Microbial test is superior to culture and PCR testing.

The MiDOG All-in-One Microbial test is superior to culture and PCR testing.

Next-Gen Sequencing has increasingly helped researchers and veterinarians characterize the skin, oral, ear, gut, and fecal microbiome of pets. This translates to direct treatment. For example, MiDOG NGS technology helped Nola, a 3-year-old Affenpinscher that was suffering from a urinary tract infection. Her veterinarian, Dr. Michael Morgan from Tustin, CA, collected her urine sample via cystocentesis and sent it out for both culture testing and to MiDOG for NGS testing. The culture results were a “no growth”, meaning the amount of bacteria growth fell below the threshold for a urinary tract infection. However, the MiDOG All-in-One Test showed a huge overgrowth of Proteus mirabilis, which is a facultatively anaerobic bacteria that the culture test missed. With this informed knowledge, Dr. Morgan was able to come up with the best treatment plan for Nola and help her recover!

The MiDOG All-in-One Microbial Test offers comprehensive diagnostic information for all different types of animals suffering from an infection. Utilizing NGS technology to detect and quantify all microbial DNA through untargeted and comprehensive sequencing and quantitative comparisons to reference databases, the MiDOG NGS technology provides a useful opportunity to shed light on the microbial makeup of your pet’s microbiome for clinical application. The MiDOG microbiome test is a microbial identification test grounded on scientific research that provides veterinarians DNA evidence for guided treatment with proven results.

 

Next-Gen Sequencing

 

Find out if your vet uses MiDOG before you book your next appointment!

References:

  1. Weir, M., Ruotsalo, K., & Tant, M. (2022). Dna Testing | VCA Animal Hospitals. Retrieved 6 May 2022, from https://vcahospitals.com/know-your-pet/dna-testing
  2. Van Borm, S., Belák, S., Freimanis, G., Fusaro, A., Granberg, F., & Höper, D. et al. (2014). Next-Generation Sequencing in Veterinary Medicine: How Can the Massive Amount of Information Arising from High-Throughput Technologies Improve Diagnosis, Control, and Management of Infectious Diseases?. Veterinary Infection Biology: Molecular Diagnostics And High-Throughput Strategies, 415-436. doi: 10.1007/978-1-4939-2004-4_30
  3. Behjati, S., & Tarpey, P. (2013). What is next generation sequencing?. Archives Of Disease In Childhood – Education &Amp; Practice Edition, 98(6), 236-238. doi: 10.1136/archdischild-2013-304340
  4. Krumbeck, J., Holden, N., & Malka, S. (2022). The Importance of Next-Generation Sequencing in Avian Veterinary Medicine – LafeberVet. Retrieved 6 May 2022, from https://lafeber.com/vet/the-importance-of-next-generation-sequencing-in-avian-veterinary-medicine/

Categories: Next-Gen DNA Sequencing Technology, Veterinarian Guides, Veterinary Best Practices, Veterinary Dermatology

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Validated by Veterinarians

“Clients expect their veterinarians to stay up to date on all matters that affect the health and well being of their non-human family members. The current technique that we have used to determine the presence and antibiotic sensitivity of organisms causing disease in our pets is over a century old.

With the emergence of dangerous antibacterial resistance, it is critical that veterinarians are able to offer laser focused diagnostics and treatment. MiDog enables us to offer care that exceeds the typical standard of care.”

Bernadine Cruz, DVM, Laguna Hills Animal Hospital Laguna Woods, CA

“I love the absolute abundance and comparing the fungal with bacterial infection. I do not worry as much about getting a false negative urinary infection reading as I do with traditional urine cultures. Several times the same urine would culture negative but MiDOG would detect pathogens.”

Michael Morgan, DVMQuail Animal Hospital, Tustin, CA

“The MiDOG All-in-One Test is amazing, I would use it instead of culture and sensitivity.  Such rapid and detailed results, I will reach for MiDOG before culture next time!

Thank you very much MiDOG, for sharing the opportunity to try your technology.”

Martha Smith-Blackmore, DVM, President of Forensic Veterinary Investigations, LLC – Boston, MA

The MiDOG All-in-One Microbial Test is our new gold standard of pathogen identification. The results are so accurate and valuable – especially with assessing both bacterial and fungal infections with the same sample.

Thank you MiDOG!”

Kathy Wentworth, DVM, Diplomate ABVP Canine and Feline Practice – PetPoint Medical Center, Irvine, CA

“The MiDOG staff was extremely helpful and supportive.”

Cathy Curtis, DVM – London, UK

“I have had great results using the MiDOG® Test. Compared to traditional culture tests, I am better able to target the treatment for dogs because the MiDOG® Test is so sensitive that it identifies all pathogens including bacteria and fungi, as well as antibiotic sensitivity.

The cost and turnaround time are about the same as a culture test, but I get much more data. The test has great performance and I believe the NGS technology will be a game changer for veterinarians treating dogs with lesions or other infections.”

Michael Kavanagh, DVM, Practice owner – Saddleback Animal Hospital, Tustin, CA

“It’s helpful to have an NGS spectrum because it gives you a broader insight of what’s happening and what might be going on.”

Richard Harvey BVSc DVD DipECVD PhD FRSB FRCVS – European Specialist in Veterinary Dermatology – Head of Dermatology, Willows Veterinary Centre & Referral Service – Solihull, England UK

“I have been using MiDog for over 4 years now and exclusively as my test of choice for all cultures for 3 years.  It is so great to submit a culture and feel confident there will be a result when it comes back, especially for urine cultures.  The reports were intimidating at first because they contain so much information.  After the first few, I am now quickly able to glance over it and pick out the highlights.  I can then come back later and pour over all the details.  I have been extremely pleased with my patients’ results using the test as well.  I don’t envision ever going back to traditional culture and susceptibilities again.”

Brian M. Urmson, DVM, Columbiana Veterinary Associates

“As an exotic veterinarian, there are numerous tests we have to consider to check specific bacterial and fungal organisms based on the species. MiDOG eliminates the need for many of these separate samples and provides definitive results quickly to help us treat our patients more efficiently and effectively. The lab is wonderful to work with and has never rejected our samples- they even processed a lizard toe we amputated and determined the cause of skin infection.”

Dr. Melissa Giese, Chicago Exotics Animal Hospital

“MiDOG’s diagnostic approach offers the unique ability to identify pathogens that evade traditional culture and sensitivity testing. I have found that adding a molecular based testing approach in the form of Next Generation Sequencing (NGS) from MiDOG to my routine diagnostic cultures can be extremely helpful in the identification and diagnosis of uncommon pathogens in veterinary medicine.”

Dr. Wayne Rosenkrantz, Animal Dermatology Clinic – Tustin

“She [Dr. Krumbeck] really did a great job of making complicated concepts accessible and demonstrating the value of your services. I’m really looking forward to working with MiDOG on my research project!”

Dr. Yaicha Peters, Animal Dermatology Clinic – San Diego

“As a proud collaborator with MiDOG, I deeply appreciate their dedication to fostering partnerships between industry and veterinary experts. Their commitment to enhancing diagnostic quality for veterinarians is commendable. In my experience, their support has been invaluable, earning them a ‘Double A+, Triple Star’ rating. Their assistance has been faultless, contributing significantly to the success of my projects and studies. I eagerly anticipate our continued collaboration.”

Dr. Richard Harvey, BVSc DVD DipECVD PhD FRSB FRCVS; European Specialist in Veterinary Dermatology

“We’re seeing that, if we prescribe too many antibiotics or they’re taken too frequently, animals are developing inappropriate or pathogenic strains of bacteria. We’re also seeing that our antibiotics are just not working against them anymore… It’s a good example of why we need better diagnostic testing, like MiDOG, so that we’re selecting the correct antibiotic every time our patients have an infection.”

Dr. Alissa Rexo, DVM, CVA, DACVD, Mid-Atlantic Veterinary Dermatology