A quick guide to Mycoplasma hyopneumoniae diagnostics

Sample types for early detection of Mycoplasma hyopneumoniae is a popular post on this blog. Dr. Pieters, head of the MycoLAb at the University of Minnesota created an online quick guide to help swine practitioners decide which sample type they should collect if they are looking for Mycoplasma hyopneumoniae.

This guide is available at http://z.umn.edu/MycoplasmaDiagnostics

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A new diagnostic test to differentiate rotavirus subtypes

In this paper published in the Journal of Veterinary Diagnostic Investigation, PhD-candidate Talita Resende from Dr. Vannucci’s lab, shares a novel diagnostic technique to detect various rotavirus species using newly developed markers.

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2016 Veterinary Diagnostic Laboratory Report: a new director, PRRS, PEDV, and Senecavirus

The Veterinary Diagnostic Laboratory’s mission is to protect and promote animal and human health through early detection and monitoring of animal diseases.
The 2016 report was published last month and we are compiling here the highlights related to swine. We can also read the full 2016 UMN VDL report.

  • In April 2016, the VDL welcomed its new director Dr. Jerry Torrison.
  • More than 50% of the procedures  in the VDL were related to the porcine species last year.
  • A new multiplex PCR test that combines Porcine Epidemic Diarrhea Virus (PEDv), Porcine Deltacoronavirus (PDCoV) and Transmissible Gastroenteritis Virus (TGEV) into one assay was implemented into the Molecular Diagnostic clinical testing schedule effective October 31st, 2016. The new assay provides clients with timely, quality results for all three viruses at the same time. The VDL ran 40,131 PEDv and PDCoV Multiplex Real Time PCR tests and 5,238 Triplex (PEDv/TGE/PDCoV) RT-PCR tests.
    Additionally, the Serology lab conducted intensive testing in collaboration with Zoetis for validation of PED antibody test kit which they are planning to release on the market soon.
  • Seneca Valley Virus PCR was validated and is part of routine testing. 3,205 Senecavirus A EZ Real time RT-PCR tests were run. An ELISA test for antibodies to Seneca Valley Virus in pigs is also available.
  • The IHC lab participated in the 2016 AAVLD/NVSL Program for Inter-laboratory Comparison, and scored 100% in its detection of Porcine Circovirus type 2 in the test samples provided.

Science Page: Continued reporting on unusual CNS cases

This is our Friday rubric: every week a new Science Page from the Bob Morrison’s Swine Health Monitoring Project. The previous editions of the science page are available on our website.

This week, the Science Page focuses on unusual Central Nervous system (CNS) cases that have been emerging in the past few years. Etiological agents like porcine teschovirus (PTV), porcine enteroviruses (PEV), porcine sapelovirus (PSV), and atypical porcine pestivirus (APPV) have been implicated in those cases leading to the creation of a set of criteria to positively identify a CNS case.

Three criteria are therefore required: identifying the clinical signs, a positive PCR test for one or more of the viruses, and histological results consistent with viral encephalitis from spinal cord or brain tissue.

Key points from this week edition:

  • An apparent increase in the number of cases associate with atypical neurological signs have been observed over the last two years.
  • Since then, the Veterinary Diagnostic Laboratories (VDLs) have identified a set of criteria required to meet the CNS case definition.
  • MSHMP will continue reporting CNS cases diagnosed at the KSU, ISU, SDSU, and UMN VDLs.

Take a look at the number of cases recorded since September 2016.

The latest trends in PRRSV diagnostics: less serum samples, more oral fluids, and more 1-7-4 RFLP pattern

Today, Dr. Albert Rovira from the University of Minnesota, Veterinary Diagnostic Laboratory shares with us the trends he has observed in PRRSV diagnostics over the past years. The findings can be found in the slideshow below.

  • The use of tissue samples follows a seasonal pattern and represents clinical cases with a percent of positives of 30%
  • The number of oral fluid samples is increasing. Used for monitoring positive farms and more recently for surveillance in negative farms as well:: 15% of positive samples
  • The number of blood swabs, serum samples, and semen samples, typically used for surveillance in negative farms, is decreasing. Lowest percent of positive samples: 8%
  • RFLP patterns are changing over time. In the past years, 1-7-4 > 1-3-4 or 1-8-4 or 1-4-4
Reminder: what is a RFLP pattern?

RFLP stands for Restriction Fragment Length Polymorphism and is a technique used to detect nucleotide changes in a genetic sequence. The genetic material is put in contact with restriction enzymes which are very specific to a genetic sequence. If the enzyme recognizes the sequence pattern, it will cleave the DNA or RNA fragment. After that, a type is determined based on the number of fragments and its size.

For example with PRRSV, three enzymes are used and the number of fragments each of them produces makes up the numbers of the RFLP pattern. Currently, the RFLP type is not actually performed in the lab. Instead, it is predicted based on the ORF5 RNA sequence and the knowledge of the cutting capabilities of each enzyme.

Therefore, the RFLP pattern gives us a way to cluster PRRSV strains in groups but very little indication about how similar they are to each other.