Srinand S, Robinson R A, Collins J E, Nagaraja K V
Department of Clinical and Population Sciences, College of Veterinary Medicine, University of Minnesota, St Paul 55108, USA.
Am J Vet Res. 1995 Sep;56(9):1163-8.
Two indirect ELISA containing outer membrane protein (OMP) and lipopolysaccharide (LPS) antigens from a field isolate of Salmonella choleraesuis var kunzendorf were developed and evaluated in experimentally infected and uninfected control pigs. Experimentally induced infection with S choleraesuis was successfully established in 10 pigs by oral inoculation with 10(8) organisms, and 3 pigs died of clinical salmonellosis at postinoculation (PI) weeks 1, 2, and 4. Swab specimens from tonsils, nostrils, and rectum of pigs were obtained for culture, and sera were evaluated at weekly intervals for 9 weeks after inoculation. The ELISA containing OMP and LPS antigens with either anti-swine IgG or protein albumin-to-globulin ratio (antiglobulin) conjugates were standardized for serologic evaluation. All 4 ELISA (2 OMP and 2 LPS) detected seroconversion by PI week 3 and had sensitivities and specificities of 97.8 and 88.8, 100 and 100, 95.6 and 88.8, and 93.3 and 72.5%, at their ideal cutoff points (negative mean optical density +2 SD). There was excellent agreement between all 4 ELISA systems as determined by kappa values. Cultures of fecal, tonsil, and nasal swab specimens were positive for S choleraesuis until the fourth week of infection. Fecal swab specimens from 1 pig were positive for S choleraesuis until PI week 7. Persistent infection after antemortem culture results were negative was detected by all 4 ELISA, which indicated consistently high titers until the end of PI week 9.(ABSTRACT TRUNCATED AT 250 WORDS)
开发了两种间接ELISA,其包含来自猪霍乱沙门氏菌kunzendorf血清型田间分离株的外膜蛋白(OMP)和脂多糖(LPS)抗原,并在实验感染和未感染的对照猪中进行了评估。通过口服接种10⁸个菌体,在10头猪中成功建立了猪霍乱沙门氏菌的实验性感染,3头猪在接种后(PI)第1、2和4周死于临床沙门氏菌病。采集猪扁桃体、鼻孔和直肠的拭子标本进行培养,并在接种后9周内每周评估血清。含有OMP和LPS抗原以及抗猪IgG或蛋白白蛋白与球蛋白比率(抗球蛋白)结合物的ELISA用于血清学评估的标准化。所有4种ELISA(2种OMP和2种LPS)在PI第3周时检测到血清转化,在其理想临界值(阴性平均光密度+2 SD)下,敏感性和特异性分别为97.8%和88.8%、100%和100%、95.6%和88.8%以及93.3%和72.5%。通过kappa值确定,所有4种ELISA系统之间具有极好的一致性。粪便、扁桃体和鼻拭子标本的培养物在感染第4周前猪霍乱沙门氏菌均为阳性。1头猪的粪便拭子标本在PI第7周前猪霍乱沙门氏菌均为阳性。所有4种ELISA均检测到生前培养结果为阴性后的持续性感染,这表明直到PI第9周结束时滴度一直很高。(摘要截断于250字)