Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae

Nielsen, Roni; Bosch, J. F. van den; Plambeck, T.; Sørensen, Vibeke; Nielsen, Jens Peter

doi:10.1016/s0378-1135(99)00157-1

Cited by 26 publications

(37 citation statements)

References 11 publications

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“…It has been reported that A. pleuropneumoniae field strains produce either ApxI, ApxII, ApxI, and ApxII or ApxII and ApxIII (40). In line with other publications (12,41), nonspecific seroconversion against ApxI was detected in serovars 3, 4, 8, and 12, against ApxII in serovars 10 and 14, and against ApxIII in serovars 1, 7, 9, 10 to 12, and 14. In this regard, it has been reported (9) that the ApxI, ApxII, and ApxIII toxins are very closely related, where ApxIII is more related to ApxI (50%) than to ApxII (41%), and ApxI and ApxII are the least related members (37%) of this toxin family.…”

Section: Discussionsupporting

confidence: 91%

“…However, ApxIV is organized differently and is genetically very distant from any of the other known RTX toxins (8). In addition, the ApxI, ApxII, and ApxIII toxins of A. pleuropneumoniae were found to be antigenically related, and a serologic correlation between serovar and secretion of exotoxin by ELISA was not established (41). This potential cross-reactivity be- (40,42,43), and therefore, additional confirmation of the expression of the respective toxins was not conducted.…”

Section: Discussionmentioning

confidence: 97%

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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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Section: Discussionsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 97%

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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“…2B). These results showed that the A. pleuropneumoniae double-mutant strain is highly attenuated and that the group infected with the mutant strain can be discriminated from the wild-type-infected group based on the ApxIIA ELISA; the antigenic relationship among A. pleuropneumoniae Apx toxins described previously (17) did not cause any false-positive reaction in the recombinant ApxII-ELISA used.…”

supporting

confidence: 61%

Construction of anActinobacillus pleuropneumoniaeSerotype 2 Prototype Live Negative-Marker Vaccine

et al. 2002

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“…While ApxIV is expressed by all A. pleuropneumoniae strains, 39 ApxI expression is limited to strains 1, 5, 9-11, and 14. 16,33 To detect strains that do not express ApxI, single-plate ELISA-2 also utilizes detection of antibodies against the surface protein Tbp2.…”

Section: Discussionmentioning

confidence: 99%

“…All samples from a pig collected after inoculation were excluded if there was no indication of seroconversion by all 4 assays utilized in this study and by the confirmatory assay(s). To confirm successful antibody production against A. suis or against A. pleuropneumoniae serovars 1, 5, 9, 10, 11, and 14, which all express the ApxI RTX toxin, 16,33 an indirect ELISA was performed as previously described 42 with the following modifications. Microtiter plates e were coated with 100 μl of the ApxI recombinant polypeptide at a concentration of 1.5 μg/ml in phosphate buffered saline (PBS, pH 7.2) per well, and the plates were incubated overnight at room temperature.…”

Section: Methodsmentioning

confidence: 99%

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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Evaluation of an indirect enzyme-linked immunosorbent assay (ELISA) for detection of antibodies to the Apx toxins of Actinobacillus pleuropneumoniae

Cited by 26 publications

References 11 publications

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

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

Construction of anActinobacillus pleuropneumoniaeSerotype 2 Prototype Live Negative-Marker Vaccine

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

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