PCR assay for specific detection of PRRSGard vaccine-like virus

 Transbound Emerg Dis

. 2021 Mar 24.

 doi: 10.1111/tbed.14084. Online ahead of print.

Development and validation of a reverse transcription real-time PCR assay for specific detection of PRRSGard vaccine-like virus

Gaurav Rawal 1, Wannarat Yim-Im 1, Fabian Chamba 2, Chad Smith 2, Jeff Okones 2, Charles Francisco 2, Jianqiang Zhang 1

Affiliations 

• PMID: 33763963
 
• DOI:10.1111/tbed.14084

Abstract

Increasing use of modified live virus (MLV) vaccines presents challenges to interpret positive results of porcine reproductive and respiratory syndrome virus (PRRSV) screening PCR that can detect both wild-type and vaccine strains. Instead, vaccine-specific PCR provides a convenient tool to detect vaccine-like virus from a sample. Here we report the development and validation of a real-time RT-PCR specific for PRRSGard® , a newly available commercial PRRSV-2 MLV vaccine. Analytical specificity, sensitivity, and diagnostic performance of PRRSGard PCR were evaluated and compared to a commercial PRRSV screening PCR (reference PCR). PRRSGard and reference PCRs did not cross-react with any of the 27 non-PRRSV swine pathogens. PRRSGard PCR did not cross-react with other PRRSV-2 vaccine viruses and 31 laboratory and field PRRSV-2 isolates representing various genetic lineages of PRRSV-2. PRRSGard and reference PCRs consistently detected up to 10-6 and 10-5 dilutions of PRRSGard vaccine virus, respectively. Based on testing serial dilutions of in vitro transcribed RNA, the 95% limit of detection of PRRSGard PCR was 16 genomic copies/reaction with CT cut-off value of 36 and 7 genomic copies/reaction with CT cut-off value of 37. Diagnostic performance of PRRSGard PCR was evaluated using 846 clinical samples (684 serum and 162 oral fluid samples). Compared to the reference screening PCR, diagnostic sensitivity, specificity, and agreement of PRRSGard PCR were 95.34%, 98.85%, and 97.52% with cut-off CT value of 36 and 98.14%, 96.56%, and 97.16% with cut-off CT value of 37. In addition, PRRSGard PCR was able to detect PRRSGard vaccine virus in a sample even with the co-presence of another PRRSV strain. In summary, in contrast to a reference screening PCR that detects both vaccine and field PRRSV strains, PRRSGard PCR provides a convenient tool to specifically detect PRRSGard vaccine-like virus and to inform PRRSV vaccination protocols.

Keywords: PRRSGard; PRRSV; Porcine reproductive and respiratory syndrome virus; Reverse transcription real-time PCR; Vaccine-like; reference screening PCR.

This article is protected by copyright. All rights reserved.

 

Review

 

. 2021 Feb 22;9(2):185.

 doi: 10.3390/vaccines9020185.

Commercial PRRS Modified-Live Virus Vaccines

Chanhee Chae 1

Affiliations 

• PMID: 33671826
 
• PMCID: PMC7926738
 
• DOI: 10.3390/vaccines9020185

Free PMC article

Abstract

Porcine reproductive and respiratory syndrome (PRRS) virus (PRRSV) presents one of the challenging viral pathogens in the global pork industry. PRRS is characterized by two distinct clinical presentations; reproductive failure in breeding animals (gilts, sows, and boars), and respiratory disease in growing pigs. PRRSV is further divided into two species: PRRSV-1 (formerly known as the European genotype 1) and PRRSV-2 (formerly known as the North American genotype 2). A PRRSV-2 modified-live virus (MLV) vaccine was first introduced in North America in 1994, and, six years later, a PRRSV-1 MLV vaccine was also introduced in Europe. Since then, MLV vaccination is the principal strategy used to control PRRSV infection. Despite the fact that MLV vaccines have shown some efficacy, they were problematic as the efficacy of vaccine was often unpredictable and depended highly on the field virus. This paper focused on the efficacy of commercially available MLV vaccines at a global level based on respiratory disease in growing pigs, and maternal and paternal reproductive failure in breeding animals.

Keywords: modified-live virus vaccine; porcine reproductive; respiratory syndrome virus.

Conflict of interest statement

The authors declare no conflict of interest.

The impact of PRRSV genotypes, established on the basis of ORF-5 sequences, on three production parameters in Ontario sow farms

 Prev Vet Med

. 2021 Feb 23;189:105312.

 doi: 10.1016/j.prevetmed.2021.105312.Online ahead of print.

The impact of porcine reproductive and respiratory syndrome virus (PRRSV) genotypes, established on the basis of ORF-5 nucleotide sequences, on three production parameters in Ontario sow farms

Dylan John Melmer 1, Terri L O'Sullivan 2, Amy Greer 2, Lori Moser 3, Davor Ojkic 4, Robert Friendship 2, Dinko Novosel 5, Zvonimir Poljak 2

Affiliations 

• PMID: 33676324
 
• DOI: 10.1016/j.prevetmed.2021.105312

Abstract

The porcine reproductive and respiratory syndrome virus (PRRSV) is an enveloped RNA virus, with high mutation rates and genetic variability; which is evident by the large number of discrete strains that co-circulate in swine populations. Veterinary practitioners frequently identify certain discrete PRRSV strains as having a higher clinical impact on production. However, with exception of a few strains, production impact is not well characterized for the majority of PRRSV variants. Predictive analytics, coupled with routine diagnostic sequencing of PRRSV, provide opportunities to study the clinical impact of discrete PRRSV strains on production. Thus, the primary objective of this research was to evaluate clinical impact of discrete PRRSV clades observed in Ontario sow farms. PRRS viruses were classified into discrete clades using Bayesian analysis of the nucleotide sequences of the ORF-5 region of the genome. Production data were gathered through veterinary clinics from herds participating in the ongoing PRRSV surveillance system. Data about pre-weaning mortality, sow mortality, and abortion rates were measured up to 8 weeks post initial PRRSV outbreak. Through conventional regression analysis, results support that clinical impact of the viruses varied among clades over time for abortion rate (p = 0.05) and pre-weaning mortality (p < 0.01). Using predictive modelling approaches based on grouped K-fold cross-validation, it was identified that PRRSV clade designations and other measured factors showed low predictive performance for abortion (R2 = 0.07), pre-weaning mortality (R2 = 0.09), and sow mortality (R2 = 0.04). Clade designation consistently showed moderate importance for abortion and pre-weaning mortality, with clade 2 viruses being identified, on average, as having higher impact. These results demonstrate that the prediction of clinical impact, through production parameters, based on phylogenetic classification of PRRS viruses is possible. However, very high impact outbreaks were difficult to predict across production parameters. More surveillance-derived data are required to continue to improve predictive performance of the models.

Keywords: BEAST; Bayesian; Ontario; PRRSV; Phylogenetics; Porcine reproductive and respiratory syndrome; Predictive modeling; Random forest; Swine.

Copyright © 2021 Elsevier B.V. All rights reserved.

 Porcine Health Manag

. 2021 Feb 26;7(1):21.

 doi: 10.1186/s40813-021-00203-4.

Effect of PRRSV stability on productive parameters in breeding herds of a swine large integrated group in Spain

D Torrents 1 2, J Miranda 3, P C Gauger 4, A Ramirez 4, Dcl Linhares 4

Affiliations 

• PMID: 33637120
 
• DOI: 10.1186/s40813-021-00203-4

Free article

Abstract

Background: In breeding herds, porcine reproductive and respiratory syndrome (PRRS) clinically manifests as increased abortions, number of stillbirths, and pre-weaning mortality, and as a direct consequence, results in a decrease of the number of piglets weaned per sow per year. Breeding farm classification according the PRRS virus (PRRSV) status (unstable or stable) is a key control strategy for this disease. The aim of this study was to evaluate the production improvement related to achieving a PRRSV stable status in breeding herds in Spain. For this purpose, epidemiological and productivity data were collected from a systematic PRRSV monitoring program in 35 breeding herds from a large integrated swine group in Spain. A comparative statistical analysis was conducted using four key production indicators (KPI) between different PRRSV status and a generalized linear mixed model: weekly abortions/1000 sows (ABTHS), born-alive rate (BAR), pre-weaning mortality rate (PWMR), and number of weaned piglets per 1000 sows (WPTHS).

Results: From the 35 monitored farms during a total period of 58 weeks, we collected 49 to 58 weeks of production data and PRRSV classification status for each study farm. This represented a total of 1997 (741 unstable and 1256 stable) weekly data collected that was eligible for the KPI comparative study. PRRSV stability was associated with significant improvement in BAR (+ 1.10 %, p < 0.001), PWMR (-0.88 %, p < 0.002) and WPTHS (+ 24.52, p < 0.0001).

Conclusions: These results demonstrate for the first time the improved production due to achieving PRRSV stability in breeding herds under field conditions in a European country. Increased number of born-alive piglets and a reduction of piglet pre-weaning mortality represents an increase of 1.28 weaned piglets per sow per year if PRRSV stability was achieved and maintained for one-year period in a breeding farm.

Keywords: Farm classification; Monitoring; Production parameters; Sows; Weaned piglets.

Antibody responses to PRRSv, IAV, and M. hyopneumoniae from weaning to the end of the finisher stage in fourteen groups of pigs in Ontario

 BMC Vet Res

. 2021 Feb 17;17(1):82.

 doi: 10.1186/s12917-021-02756-6.

Antibody responses to porcine reproductive and respiratory syndrome virus, influenza A virus, and Mycoplasma hyopneumoniae from weaning to the end of the finisher stage in fourteen groups of pigs in Ontario, Canada

Elana Raaphorst 1, Abdolvahab Farzan 2 3, Robert M Friendship 4, Brandon N Lillie 1

Affiliations 

• PMID: 33596907
 
• DOI: 10.1186/s12917-021-02756-6

Abstract

Background: Respiratory diseases are among the most important factors affecting swine farm productivity in Canada. The objectives of this study were to investigate antibody responses to porcine reproductive and respiratory syndrome virus (PRRSV), influenza A virus (IAV), and Mycoplasma hyopneumoniae (M. hyopneumoniae) from weaning to the end of the finisher stage on a subset of commercial swine farms in Ontario, Canada, and to examine the association between nursery diet and antibody responses.

Results: Overall, older pigs were more likely to test seropositive for PRRSV and less likely to test seropositive for M. hyopneumoniae (p < 0.001). Pigs were more likely to test seropositive for IAV at weaning and the end of the grower and finisher stages compared to the end of nursery (p < 0.001). Pigs that were seropositive for IAV were more likely to test seropositive for both PRRSV and M. hyopneumoniae (p < 0.001). Two, 9, and 4 groups that had more than 20% of pigs seropositive to PRRSV, IAV, and M. hyopneumoniae, respectively, from the end of nursery to the end of finisher were classified as seropositive. Pigs fed a plant-based (low complexity) diet during nursery were more likely to be seropositive for PRRSV (p < 0.001) but there were no significant differences in seropositivity to IAV or M. hyopneumoniae due to nursery diet complexity.

Conclusions: This study provides information regarding changes in serum antibody in pigs across different stages of production and highlights periods of vulnerability. Additionally, these findings may encourage further research into the effects of nursery diet complexity on disease susceptibility and immune response.

Keywords: Antibody response; ELISA; Influenza A virus; Mycoplasma hyopneumoniae; Nursery diet; Porcine reproductive and respiratory syndrome; swine.

 Prev Vet Med

. 2021 Jan 27;189:105270.

 doi: 10.1016/j.prevetmed.2021.105270.Online ahead of print.

Association of wild-type PRRSV detection patterns with mortality of MLV-vaccinated growing pig groups

Cesar A A Moura 1, Reid Philips 2, Gustavo S Silva 1, Alejandro Ramirez 1, Phillip C Gauger 1, Derald J Holtkamp 1, Daniel C L Linhares 3

Affiliations 

• PMID: 33550121
 
• DOI: 10.1016/j.prevetmed.2021.105270

Abstract

PRRS is a viral disease characterized by increasing reproduction losses in breeding herds and worsening performance of growing pigs, which leads to a considerable economic impact. A better understanding of the differences in the infection patterns of the virus in growing pig batches would help to develop cost-effective surveillance methods and disease control and elimination programs. Therefore, field studies documenting growing pig productivity according to the patterns of wild-type-PRRSV (wt-PRRSV) detection in the field are needed. This study was conducted with the objectives to (1) characterize patterns of wt-PRRSV-1 and wt-PRRSV-2 RNA detection over time in modified-live virus (MLV)-vaccinated batches of growing pigs raised in pig-dense regions of the USA; (2) compare wean-to-finish mortality among batches of growing pigs characterized with the different patterns of wt-PRRSV-1 and wt-PRRSV-2 detection; and (3) compare wean-to-finish mortality among batches of growing pigs characterized with the different patterns of wt-PRRSV-2 detection and vaccinated with two different doses of PRRS MLV vaccine. Eighty-one batches of growing pigs were originated from PRRSV positive-stable and unstable sow farms and vaccinated with two different doses of PRRS MLV vaccine. All batches were monitored for wt-PRRSV by testing six oral fluids every three weeks from weaning to marketing. Diagnostics were conducted to detect wt-PRRSV-1 and wt-PRRSV-2 by RT-qPCR testing and ORF-5 region sequencing. K-means clustering analysis was applied to identify batches sharing similar patterns of wt-PRRSV detection over time. Regression analyses were used to compare mortality among batches of growing pigs characterized with different patterns of wt-PRRSV detection over time. Thirty-eight percent of batches were detected with wt-PRRSV-1 during the growth phase, with three different patterns of detection. Detection of wt-PRRSV-1 was not associated with significant increase on mortality. Ninety-one percent of batches were detected with wt-PRRSV-2 during the growth phase, with four different patterns of detection. Batches originated from PRRSV positive-unstable farms had highest mortality rate (p < 0.0001) and were characterized as Unstable wt-PRRSV-2 detection pattern. Batches characterized with Early wt-PRRSV-2 detection pattern had higher mortality than batches characterized by Mid, Late and No wt-PRRSV-2 detection during the growth phase (p < 0.0001). Batches with Mid wt-PRRSV-2 detection had higher mortality than batches characterized with Late wt-PRRSV-2 detection (p < 0.0124). Mortality rate of batches characterized with Unstable and Early wt-PRRSV-2 detection patterns was lower when pigs were vaccinated with two doses of PRRS MLV vaccine, compared to batches that received only one dose. Results presented in this study suggested that early wt-PRRSV exposure on pig populations was associated with higher wean-to-finish mortality. Additionally, results suggested that vaccination with two PRRS MLV doses was associated with lower mortality rate, when growing pig populations had early wt-PRRSV exposure.

Keywords: Field epidemiology; Growing pigs; Mortality; PRRSV; Swine.

Copyright © 2021 Elsevier B.V. All rights reserved.