Free accessResearch articleFirst published online 1993-7
Development and Evaluation of a Mixed-Antigen ELISA for Serodiagnosis of Actinobacillus Pleuropneumoniae Serotypes 1,5,and 7 Infections in Commercial Swine Herds
A mixed-antigen enzyme-linked immunosorbent assay (ELISA) containing serotype-specific polysaccharide antigens from serotypes 1, 5, and 7 of Actinobacillus pleuropneumoniae was developed. This ELISA was evaluated using sera from experimentally infected pigs. With a negative cutoff value of 0.250 (optical density at 405 nm), sensitivity and specificity were determined to be 96% and 99.5%, respectively. The ELISA was further evaluated using sera from commercial swine. Sows and suckling piglets were found to be the best age groups for detection of positive reactors to A. pleuropneumoniae. The mixed-antigen ELISA could be used in a herd health monitoring program for detection of A. pleuropneumoniae infections.
References
1.
AltmanEBrissonJRPerryMB: 1986, Structural studies of the capsular polysaccharide from Haemophilus pleuropneumoniae serotype 1. Biochem Cell Biol64:707–716.
2.
BeynonLMPerryMBRichardsJC: 1991, Structure of the capsular polysaccharide from Actinobacillus pleuropneumoniae serotype 7. Carbohydr Res209:211–223.
3.
BosséJTJohnsonRPRosendalS: 1990, Capsular polysaccharide antigens for detection of serotype specific antibodies to Actinobacillus pleuropneumoniae. Can J Vet Res54:320–325.
4.
BosséJTJohnsonRPRosendalS: 1990, Serodiagnosis of pleuropneumonia using enzyme-linked immunosorbent assay with capsular polysaccharide antigens of Actinobacillus pleuropneumoniae serotypes 1, 2, 5 and 7. Can J Vet Res54:427–431.
5.
BradfordM: 1976, A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Anal Biochem95:248–254.
6.
DuboisMGillesKAHamiltonJK: 1956, Calorimetric method for determination of sugars and related substances. Anal Chem28:350–356.
7.
GardnerIABosséJTSheldrakeRF: 1991, Serological response to Actinobacillus pleuropneumoniae serovar 7 infection in a commercial pig herd. Aust Vet J68:349–352.
8.
GilbrideKARosendalS: Evaluation of a selective medium for isolation of Haemophilus pleuropneumoniae. Can J Comp Med47:445–450.
9.
GunnarssonA: 1979, Serologic studies on porcine strains of Haemophilus parahaemolyticus (pleuropneumoniae): Extraction of typespecific antigens. Am J Vet Res39:469–472.
10.
NicoletJParozPHKrawinklerMBaumgartnerA: 1981, An enzyme-linked immunosorbent assay, using an EDTA-extracted antigen for the serology of Haemophilus pleuropneumoniae. Am J Vet Res42:2139–2142.
11.
NielsenRThomsenADDuedahlVesterlund S: 1976, Pleuropneumonia caused by Haemophilus parahaemolyticus: An attempt to control the disease at two progeny research stations by serological blood testing followed by removal of seropositive animals and their littermates. Nord Vet Med28:349–352.
12.
O'ReillyTNivenDF: 1986, Tryptone-yeast extract as a culture medium for Haemophilus pleuropneumoniae and Haemophilus parasuis to be used as challenge inocula. Can J Vet Res50:441–443.
13.
RosendalSMitchellWR: 1983, Epidemiology of Haemophilus pleuropneumoniae infection in pigs: A survey of Ontario pork producers, 1981. Can J Comp Med47:1–5.
14.
RosendalSMittalKR: 1985, Serological cross-reactivity between a porcine Actinobacillus strain and Haemophilus pleuropneumoniae. Can J Comp Med49:164–170.
15.
SebunyaTNKSaundersJROsborneAD: 1983, Dose response relationship of Haemophilus pleuropneumoniae aerosols in pigs. Can J Comp Med47:54–56.
16.
VaillancourtJPMartinaeuGPLariviereS: 1988, Serological follow-up in breeding herds infected with Actinobacillus pleuropneumoniae serotype 1 using the tube agglutination test with 2-mercaptoethanol. Prev Vet Med6:263–274.
17.
WillsonPJFalkGKlashinskyS: Detection of Actinobacillus pleuropneumoniae infection in pigs. Can Vet J28:111–116.
