Identification of a genetically diverse sequence of porcine reproductive and respiratory syndrome virus in Slovenia and the impact on the sensitivity of four molecular tests

Toplak, Ivan; Rihtarič, Danijela; Hostnik, Peter; Grom, Jože; Štukelj, Marina; Valenčak, Z.

doi:10.1016/j.jviromet.2011.09.019

Cited by 25 publications

(22 citation statements)

References 21 publications

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“…Partial depopulation and the test and removal strategy can decrease the time needed for the elimination of PRRSV from a herd. The previous observation for Slovenia was that PRRSV was endemically present in swine across the whole territory as a result of low biosecurity standards, intensive trading and importation of pigs from different countries (Toplak et al, 2012). According to the results of the RT-PCR method and direct sequencing of six PRRSV-positive samples, we confirmed that during the entire period of study the same strain of PRRSV was present, and that herd closure and other preven-…”

Section: Discussionsupporting

confidence: 79%

“…The results of tested samples were divided into four groups; samples with an S/P lower than 0.4 (negative), samples with an S/P between 0.4 and 1 (low positive), samples with an S/P between 1 and 2 (positive), and samples with an S/P higher than 2 (high positive). Table 2 Results of ELISA for the detection of PRRS antibodies (Donadeu et al, 1999;Toplak et al, 2012). The PRRSV-negative serum samples and the Lelystad virus (Type 1) were used as negative and positive controls, respectively.…”

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

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Serum inoculation as a possibility for elimination of porcine reproductive and respiratory syndrome (PRRS) from a farrow-to-finish pig farm

Štukelj

Plut

Toplak

2015

Acta Veterinaria Hungarica

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The large heterogeneity among porcine reproductive and respiratory syndrome virus (PRRSV) isolates is probably the main obstacle to its effective control using current commercial vaccines. Intentionally exposing all breeding pigs to PRRSV circulating on the farm could eliminate porcine reproductive and respiratory syndrome (PRRS) from the herd. The objective of this study was to eliminate PRRS from a farrow-to-finish pig farm by serum inoculation. The owner was acquainted with the strict biosecurity measures. Breeding pigs were immunised with serum, which was obtained from PRRSV-positive weaners from the same farm. The percent of antibody high positive breeding pigs decreased six months after serum inoculation, while 34 months after serum inoculation no more antibody high positive pigs were detected and 56.8% of breeding pigs and all other categories were free of antibodies. In the breeding herd no virus was detected during all testing while PRRSV circulated in 2-month-old weaners until 12 months after serum inoculation. Later all tested samples from weaners, growers and fatteners were negative. Herd closure and the adoption of strict biosecurity measures are essential. Serum inoculation of the breeding herd proved to be a successful measure for eliminating PRRS from this farrow-to-finish farm.

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Section: Discussionsupporting

confidence: 79%

Section: Enzyme-linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Serum inoculation as a possibility for elimination of porcine reproductive and respiratory syndrome (PRRS) from a farrow-to-finish pig farm

Štukelj

Plut

Toplak

2015

Acta Veterinaria Hungarica

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“…However, the high mutation rate, rapid evolution, and genetic variability of PRRSV strains complicate the development of long-term reliable diagnostic assays, and consequently cases of false-negative results with commercially available assays have been reported (3)(4)(5).…”

mentioning

confidence: 99%

Comparison of Commercial Real-Time Reverse Transcription-PCR Assays for Reliable, Early, and Rapid Detection of Heterologous Strains of Porcine Reproductive and Respiratory Syndrome Virus in Experimentally Infected or Noninfected Boars by Use of Different Sample Types

et al. 2013

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The aims of this study were to compare three commercial porcine reproductive and respiratory syndrome virus (PRRSV) realtime reverse transcription-PCR (RT-PCR) assays for detection of genetically diverse PRRSV isolates in serum, semen, blood swabs, and oral fluids collected from experimentally infected boars and to evaluate the effects of sample pooling. Six groups of three boars negative for PRRSV were each inoculated with one of six PRRSV isolates (sharing 55 to 99% nucleotide sequence identity in ORF5). Samples were collected on days ؊2, P orcine reproductive and respiratory syndrome virus (PRRSV) continues to be the most important pathogen affecting pigs in North America (1). PRRSV is a small, enveloped, single-stranded positive-sense RNA virus of the family Arteriviridae and can be divided into two genotypes: type 1 (European type [EU]) and type 2 (North American type [NA]) (2). Although a variety of commercial vaccines are available on the global market, the virus remains difficult to control, and the demand for PRRSV-naïve replacement genetics and, with it, the need for highly sensitive and specific assays that can detect genetically diverse strains and provide information on the most appropriate samples for testing continue to grow.Presently, many boar studs in the United States are PRRSV negative and are routinely tested for PRRSV to ensure that PRRSV-free semen is used in breeding herds for artificial insemination (1). If previously negative boar studs become infected with PRRSV, it is critical to detect the virus as soon as possible so that any shipments of possible PRRSV-contaminated semen can be stopped.The reverse transcription-PCR (RT-PCR) method in the realtime format is one of the most commonly used techniques for detection of PRRSV RNA because of its sensitivity and specificity and relatively short test turnaround time. However, the high mutation rate, rapid evolution, and genetic variability of PRRSV strains complicate the development of long-term reliable diagnostic assays, and consequently cases of false-negative results with commercially available assays have been reported (3-5).Active PRRSV surveillance in boar studs relies mainly on collection and testing of serum, semen, blood swabs, and, more recently, oral fluids (6, 7). The choice of sample type should take into consideration the availability and ease of collection in addition to the sensitivity and specificity of the RT-PCR assay. Studies have shown that PRRSV RNA can be detected in boar serum, oral fluids, and blood swabs as early as 24 to 48 h postinfection and in semen samples as early as 48 to 120 h postinfection (6-9). Isolatespecific differences in the levels of PRRSV replication and shedding in the host have been reported (10), and as veterinarians and diagnosticians consider using alternative sampling methods, it is important to conduct an unbiased comparison of the ability of different commercial real-time RT-PCR tests to detect genetically diverse isolates of PRRSV in new and conventional sample types.The sample collection ...

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“…Toplak et al demonstrated that the delay to diagnosis of MWS has decreased significantly over the past 40 years (29). These authors suggest that increasing knowledge and awareness plus availability of genetic testing led to this progress (29). Our study determined that other factors, such as distance to the facility of MWS expertise and socioeconomic status of the patient, did not impact on time to diagnosis.…”

Section: Discussionmentioning

confidence: 55%

“…While the age of MWS diagnosis was not specified, the overall age at onset of symptoms for the entire CAPS series was Ͻ10 years in 86% of patients. Toplak et al demonstrated that the delay to diagnosis of MWS has decreased significantly over the past 40 years (29). These authors suggest that increasing knowledge and awareness plus availability of genetic testing led to this progress (29).…”

Section: Discussionmentioning

confidence: 99%

Challenges in Diagnosing Muckle‐Wells Syndrome: Identifying Two Distinct Phenotypes

Kuemmerle‐Deschner

Samba

Tyrrell

et al. 2014

Arthritis Care & Research

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Objective. The diagnosis of Muckle-Wells syndrome (MWS) remains challenging due to the clinical heterogeneity and lack of diagnostic criteria. The aims of this study were to describe key elements of the diagnostic evaluation process in MWS and compare identified variables between patients diagnosed in childhood and adulthood. Methods. A cohort study of consecutive patients with a clinical and genetic diagnosis of MWS was conducted at 2 reference centers for autoinflammatory diseases. Demographic information, clinical presentation, access to care, and preclinical evaluation variables were captured. Presenting symptoms were compared between groups of patients diagnosed in childhood and adulthood. Prediction analysis explored variables associated with late diagnosis. Correspondence analysis identified clinical phenotypes. Results. A total of 34 MWS patients were included (16 males, 18 females) and median age at diagnosis was 31.5 years (range 0.5-75 years). Patients diagnosed during childhood reported musculoskeletal symptoms (62%), rash (62%), fever (54%), and abdominal pain (31%). Those diagnosed as adults described musculoskeletal symptoms (86%), rash (67%), hearing loss (52%), and fatigue (29%). Hearing loss was associated with late diagnosis, while access-to-care variables were not predictive. Correspondence analysis identified distinct clinical phenotypes as follows: an "inflammatory phenotype" (most commonly seen in patients diagnosed in childhood and characterized by relapsing fever and abdominal pain), an intermediate phenotype, and an "organ-disease" phenotype in patients diagnosed during adulthood and characterized by fatigue and hearing loss. Conclusion. Distinct clinical phenotypes were identified in patients with MWS. These are closely related to age at diagnosis. The presence of these phenotypes has to be considered when developing diagnostic criteria for MWS.

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Identification of a genetically diverse sequence of porcine reproductive and respiratory syndrome virus in Slovenia and the impact on the sensitivity of four molecular tests

Cited by 25 publications

References 21 publications

Serum inoculation as a possibility for elimination of porcine reproductive and respiratory syndrome (PRRS) from a farrow-to-finish pig farm

Serum inoculation as a possibility for elimination of porcine reproductive and respiratory syndrome (PRRS) from a farrow-to-finish pig farm

Comparison of Commercial Real-Time Reverse Transcription-PCR Assays for Reliable, Early, and Rapid Detection of Heterologous Strains of Porcine Reproductive and Respiratory Syndrome Virus in Experimentally Infected or Noninfected Boars by Use of Different Sample Types

Challenges in Diagnosing Muckle‐Wells Syndrome: Identifying Two Distinct Phenotypes

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