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.

Continue reading “A new diagnostic test to differentiate rotavirus subtypes”

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.

Detecting Senecavirus A in tissues: development of a new diagnostic test at the University of Minnesota

Summary

In Canada and the USA alike, Senecavirus A is a challenge for producers and veterinarians because of its clinical similarity to Food and Mouth Disease (FMD). Indeed, Senecavirus A, is a causative agent of swine vesicular disease with lesions developing on the snout, around the mouth and on the coronary band of the feet. Therefore, being able to differentiate Senecavirus A infections from FMD rapidly is of utmost importance to be able to take the appropriate measures.

In the past months, several diagnostic tests have been developed at the University of Minnesota to detect antibodies against Senecavirus A. The difference between those tests and the in situ hybridization (ISH) described here is that ISH targets the genetic material included in the viral particle and marks it as a red spot as can be seen on the figure below. This advantage of this method is to be able to locate the virus and gives additional information to researcher wanting to study the behavior of Senecavirus A in the body of the pig.

in situ hybridization senecavirus A pigs
Red dots and clusters represent the presence of SVV mRNA within an erosive lesion on the tongue of a pig © 2017 Resende et al.

Abstract

Seneca Valley virus (SVV) is the causative agent of an emerging vesicular disease in swine, which is clinically indistinguishable from other vesicular diseases such as foot-and-mouth disease. In addition, SVV has been associated with neonatal mortality in piglets. While a commercial SVV qRT-PCR is available, commercial antibodies are lacking to diagnose SVV infections by immunohistochemistry (IHC). Thus, a novel in situ hybridization technique—RNAscope (ISH) was developed to detect SVVRNA in infected tissues. From a total of 78 samples evaluated, 30 were positive by qRT-PCR and ISH-RNA, including vesicular lesions of affected sows, ulcerative lesions in the tongue of piglets and various other tissues with no evidence of histological lesions. Nineteen samples were negative for SVV by qRT-PCR and ISH-RNA. The Ct values of the qRT-PCR from ISH-RNA positive tissues varied from 12.0 to 32.6 (5.12 x 106 to 5.31 RNA copies/g, respectively). The ISH-RNA technique is an important tool in diagnosing and investigating the pathogenesis of SVV and other emerging pathogens.

Link to the full article