Evaluation of Actinobacillus pleuropneumoniae diagnostic tests using samples derived from experimentally infected pigs

Costa, G.; Oliveira, Simone; Torrison, Jerry; Dee, S.

doi:10.1016/j.vetmic.2010.08.023

Cited by 25 publications

(33 citation statements)

References 94 publications

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“…Specifically, pigs infected with serovars 5 and 12 did not seroconvert, and polymerase chain reaction performed on tonsils at termination of that earlier study was negative. 7 Therefore, it is possible that the animals in the present study remained uninfected perhaps due to a lower dose than required to induce infection with these particular serovars, explaining the lack of detection of antibodies. In the present study, successful infection and lack of interference by the antimicrobial treatment was confirmed by assays targeting antibodies against ApxI and ApxII or serotype-specific assays.…”

Section: Discussionmentioning

confidence: 81%

Evaluation of diagnostic assays for the serological detection ofActinobacillus pleuropneumoniaeon samples of known or unknown exposure

et al. 2013

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Abstract. Accurate diagnosis of exposure to Actinobacillus pleuropneumoniae is important for maintaining negative farms. In the present study, the ability of a dual-plate complement fixation (CF) assay and 3 commercially available enzymelinked immunosorbent assays (ELISAs; quad-plate ELISA-1, single-plate ELISA-2, and single-plate ELISA-3) in detecting serological evidence of A. pleuropneumoniae exposure was compared using serum samples of experimentally infected or vaccinated pigs, or field samples from the United States. Forty-two pigs were divided into groups of 2 pigs and were inoculated with 1 of 15 A. pleuropneumoniae strains representing all known serovars of A. pleuropneumoniae, or with Actinobacillus suis, or were vaccinated with a bacterin containing A. pleuropneumoniae serovar 1, 3, 5, or 7. Serum samples collected at the day of inoculation or vaccination and 7, 14, 21, and 28 days later were used to compare the assays. On samples from experimentally infected pigs, the dual-plate CF assay, quad-plate ELISA-1, single-plate ELISA-2, and single-plate ELISA-3 had sensitivities of 0.46, 0.74, 0.13, and 0.13 and specificities of 0.90, 1.0, 1.0, and 1.0, respectively. Vaccinated pigs were identified only by the dual-plate CF assay and the quad-plate ELISA-1. In addition, 90 serum samples with unknown A. pleuropneumoniae exposure collected under field conditions were tested with all assays. The agreement of the 4 assays on field samples was slight to fair. While several assays are available for demonstration of A. pleuropneumoniae exposure, differences in assay targets complicate test choices. Decisions on which assay or combination of assays to use depend on the specific reasons for running the assays.

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Section: Discussionmentioning

confidence: 81%

Evaluation of diagnostic assays for the serological detection ofActinobacillus pleuropneumoniaeon samples of known or unknown exposure

et al. 2013

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“…In addition, optimised PCR conditions have been developed for specific respiratory pathogens including PRRSV [22, 23], SIV [24, 25] and PCV2 [26, 27]. Most focus has been on viral rather than bacterial pathogen detection in OF but the knowledge base is increasing for Actinobacillus pleuropneumoniae , Haemophilus parasuis [28] and M. hyo [29–31]. In addition, the OF platform relies on a pooled sample at pen-level from animals that may or may not interact individually with the sampling rope and may themselves be shedding pathogen, or have antibody titres, at differing levels thereby raising potential constraints related to sensitivity of the test.…”

Section: Introductionmentioning

confidence: 99%

The use of oral fluids to monitor key pathogens in porcine respiratory disease complex

et al. 2017

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BackgroundThe usefulness of oral fluid (OF) sampling for surveillance of infections in pig populations is already accepted but its value as a tool to support investigations of porcine respiratory disease complex (PRDC) has been less well studied. This study set out to describe detection patterns of porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), swine influenza virus type A (SIV) and Mycoplasma hyopneumoniae (M. hyo) among farms showing differing severity of PRDC.The study included six wean-to-finish pig batches from farms with historical occurrence of respiratory disease. OF samples were collected from six pens every two weeks from the 5th to the 21st week of age and tested by real time PCR for presence of PRRSV, SIV and M. hyo and by quantitative real time PCR for PCV2. Data was evaluated alongside clinical and post-mortem observations, mortality rate, slaughter pathology, histopathology, and immunohistochemistry testing data for PCV2 antigen where available.ResultsPRRSV and M. hyo were detectable in OF but with inconsistency between pens at the same sampling time and within pens over sequential sampling times. Detection of SIV in clinical and subclinical cases showed good consistency between pens at the same sampling time point with detection possible for periods of 2–4 weeks. Quantitative testing of OF for PCV2 indicated different patterns and levels of detection between farms unaffected or affected by porcine circovirus diseases (PCVD). There was good correlation of PCR results for multiple samples collected from the same pen but no associations were found between prevalence of positive test results and pen location in the building or sex of pigs.ConclusionsDetection patterns for PRRSV, SIV and M. hyo supported the effectiveness of OF testing as an additional tool for diagnostic investigation of PRDC but emphasised the importance of sampling from multiple pens and on multiple occasions. Preliminary evidence supported the measurement of PCV2 load in pooled OF as a tool for prediction of clinical or subclinical PCVD at farm level.

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“…A. pleuropneumoniae can infect pigs of all ages, but clinical disease is mainly seen in growing pigs Ͼ12 weeks of age (2,3). The disease is transmitted by the aerosol route or by direct contact and is often highly contagious.…”

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confidence: 99%

“…Among the different ELISAs, those targeting ApxIV toxin to quantify circulating levels of the antibody to A. pleuropneumoniae (20), those targeting ApxI, ApxII, or ApxIII for the detection of specific antibodies to Apx exotoxins (21), those targeting capsular polysaccharides of A. pleuropneumoniae to identify antibodies against groups of its serovars (22), and serovar-specific ELISAs based on long-chain lipopolysaccharides (23) have been widely used and offer better sensitivities and specificities than the CFT. It has also been demonstrated that ELISAs utilizing different antigens or that are performed under different assay conditions may have conflicting results and cross-reactions between A. pleuropneumoniae serovars and other bacterial species, and this may be problematic (2,3). The test specificity greatly varies depending on the serologic assay performed.…”

mentioning

confidence: 99%

Simultaneous Detection of Antibodies against Apx Toxins ApxI, ApxII, ApxIII, and ApxIV in Pigs with Known and Unknown Actinobacillus pleuropneumoniae Exposure Using a Multiplexing Liquid Array Platform

Giménez-Lirola¹,

Jiang²,

Sun³

et al. 2013

Clin. Vaccine Immunol.

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c Surveillance for the presence of Actinobacillus pleuropneumoniae infection in a population plays a central role in controlling the disease. In this study, a 4-plex fluorescent microbead-based immunoassay (FMIA), developed for the simultaneous detection of IgG antibodies to repeat-in-toxin (RTX) toxins (ApxI, ApxII, ApxIII, and ApxIV) of A. pleuropneumoniae, was evaluated using (i) blood serum samples from pigs experimentally infected with each of the 15 known A. pleuropneumoniae serovars or with Actinobacillus suis, (ii) blood serum samples from pigs vaccinated with a bacterin containing A. pleuropneumoniae serovar 1, 3, 5, or 7, and (iii) blood serum samples from pigs with an unknown A. pleuropneumoniae exposure status. The results were compared to those obtained in a previous study where a dual-plate complement fixation test (CFT) and three commercially available enzyme-linked immunosorbent assays (ELISAs) were conducted on the same sample set. On samples from experimentally infected pigs, the 4-plex Apx FMIA detected specific seroconversion to Apx toxins as early as 7 days postinfection in a total of 29 pigs inoculated with 14 of the 15 A. pleuropneumoniae serovars. Seroconversion to ApxII and ApxIII was detected by FMIA in pigs inoculated with A. suis. The vaccinated pigs showed poor humoral responses against ApxI, ApxII, ApxIII, and ApxIV. In the field samples, the humoral response to ApxIV and the A. pleuropneumoniae seroprevalence increased with age. This novel FMIA (with a sensitivity of 82.7% and a specificity of 100% for the anti-ApxIV antibody) was found to be more sensitive and accurate than current tests (sensitivities, 9.5 to 56%; specificity, 100%) and is potentially an improved tool for the surveillance of disease and for monitoring vaccination compliance.

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Evaluation of Actinobacillus pleuropneumoniae diagnostic tests using samples derived from experimentally infected pigs

Cited by 25 publications

References 94 publications

Evaluation of diagnostic assays for the serological detection ofActinobacillus pleuropneumoniaeon samples of known or unknown exposure

Evaluation of diagnostic assays for the serological detection ofActinobacillus pleuropneumoniaeon samples of known or unknown exposure

The use of oral fluids to monitor key pathogens in porcine respiratory disease complex

Simultaneous Detection of Antibodies against Apx Toxins ApxI, ApxII, ApxIII, and ApxIV in Pigs with Known and Unknown Actinobacillus pleuropneumoniae Exposure Using a Multiplexing Liquid Array Platform

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