Unidad de Sanidad Animal, Centro de Investigación y Tecnología Agroalimentaria (CITA) del Gobierno de Aragón, Av. Montañana 930, 50059 Zaragoza, Spain.
Dpt. Patología Animal, Facultad de Veterinaria, Universidad de Zaragoza, 50013 Zaragoza, Spain.
Vet Microbiol. 2014 Jan 10;168(1):161-8. doi: 10.1016/j.vetmic.2013.10.024. Epub 2013 Nov 11.
Swine brucellosis by Brucella suis biovar 2 is an emerging disease whose control is based on serological testing and culling. However, current serological tests detect antibodies to the O-polysaccharide (O/PS) moiety of Brucella smooth lipopolysaccharide (S-LPS), and thus lack specificity when infections by Yersinia enterocolitica O:9 and other gram-negative bacteria carrying cross-reacting O/PS occur. The skin test with the protein-rich brucellin extract obtained from rough B. melitensis B115 is assumed to be specific for discriminating these false positive serological reactions (FPSR). However, B115 strain, although unable to synthesize S-LPS, accumulates O/PS internally, which could cause diagnostic problems. Since the brucellin skin test has been seldom used in pigs and FPSR are common in these animals, we assessed its performance using cytosoluble protein extracts obtained from B. abortus rough mutants in manBcore or per genes (critical for O/PS biosynthesis) and B. melitensis B115. The diagnostic sensitivity and specificity were determined in B. suis biovar 2 culture positive and brucellosis free sows, and apparent prevalence in sows of unknown individual bacteriological and serological status belonging to B. suis biovar 2 naturally infected herds. Moreover, the specificity in discriminating brucellosis from FPSR was assessed in brucellosis free boars showing FPSR. The skin test with B. abortus ΔmanBcore and B. melitensis B115 allergens performed similarly, and the former one resulted in 100% specificity when testing animals showing FPSR in indirect ELISA, Rose Bengal and complement fixation serological tests. We conclude that O/PS-free genetically defined mutants represent an appropriate alternative to obtain Brucella protein extracts for diagnosing swine brucellosis.
猪布鲁氏菌病由 2 型布氏菌引起,是一种新出现的疾病,其控制基于血清学检测和淘汰。然而,目前的血清学检测方法检测到布鲁氏菌光滑脂多糖(S-LPS)的 O-多糖(O/PS)部分的抗体,因此当感染携带交叉反应 O/PS 的肠沙门氏菌 O:9 和其他革兰氏阴性菌时缺乏特异性。用从粗糙型 B. melitensis B115 中提取的富含蛋白质的布鲁氏菌素提取物进行皮肤试验,被认为是特异性的,可用于区分这些假阳性血清学反应(FPSR)。然而,B115 菌株虽然不能合成 S-LPS,但会在内部积累 O/PS,这可能会导致诊断问题。由于布鲁氏菌素皮肤试验在猪中很少使用,并且 FPSR 在这些动物中很常见,因此我们使用 manBcore 或 per 基因(O/PS 生物合成关键基因)缺失的粗糙突变株和 B. melitensis B115 从流产布氏杆菌中提取的可溶蛋白提取物来评估其性能。在 2 型布氏菌培养阳性且无布鲁氏菌病的母猪以及来自自然感染 2 型布氏菌的未知个体细菌学和血清学状态的母猪中,确定了诊断敏感性和特异性。此外,还在表现出 FPSR 的无布鲁氏菌病公猪中评估了区分布鲁氏菌病和 FPSR 的特异性。用 B. abortus ΔmanBcore 和 B. melitensis B115 变应原进行皮肤试验的效果相似,当用间接 ELISA、虎红平板和补体结合血清学试验检测到 FPSR 时,前者的特异性为 100%。我们得出结论,无 O/PS 的遗传定义突变体是获得用于诊断猪布鲁氏菌病的布鲁氏菌蛋白提取物的合适替代方法。