Rafiee M, Bara M, Stephens C P, Blackall P J
Australasian Pig Institute, Queensland Department of Primary Industries, Animal Research Institute, Yeerongpilly, Queensland 4105.
Aust Vet J. 2000 Dec;78(12):846-9. doi: 10.1111/j.1751-0813.2000.tb10507.x.
To validate a polymerase chain reaction (PCR) based method. Enterobacterial Repetitive Intergenic Consensus-PCR (ERIC-PCR), for the fingerprinting of Haemophilus parasuis strains and to use that method to differentiate isolates from apparently related outbreaks of Glässer's disease on three pig farms.
ERIC-PCR was evaluated by comparing 15 different strains that represented all 15 recognised serovars in the Kielstein-Rapp-Gabrielson (KRG) scheme for serotyping H parasuis. Next, ERIC-PCR was used to examine 14 Australian field isolates of H parasuis; 12 collected from three farms suffering apparently related outbreaks of Glässer's disease and two from two other farms with no known connection.
The 15 serovar reference strains all gave unique, reproducible ERIC-PCR fingerprints. The 12 isolates from the three apparently related outbreaks all gave a single fingerprint, which was distinct from any seen in the 15 serovar reference strains and the two other Australian field isolates in the studied farms. The confirmation that all 12 isolates were the same strain allowed the development of a prevention and control program that has prevented the emergence of any further outbreaks of Glässer's disease on the three farms.
ERIC-PCR is a suitable technique for the differentiation of unrelated strains of H parasuis. The finding that the 12 field isolates of H parasuis all shared the same fingerprint is strong evidence that there was a common source of infection on all three farms. This study has shown, for the first time, that ERIC-PCR is a suitable technique for the sub-typing of H parasuis and useful for studying the epidemiology of outbreaks of Glässer's disease.
验证一种基于聚合酶链反应(PCR)的方法——肠杆菌基因间重复共有序列PCR(ERIC-PCR),用于副猪嗜血杆菌菌株的指纹图谱分析,并使用该方法区分三个猪场疑似相关的格氏病暴发中的分离株。
通过比较15种不同菌株对ERIC-PCR进行评估,这些菌株代表了基尔施泰因-拉普-加布里埃尔森(KRG)副猪嗜血杆菌血清分型方案中所有15种公认的血清型。接下来,使用ERIC-PCR检测14株澳大利亚副猪嗜血杆菌田间分离株;其中12株从三个疑似相关格氏病暴发的猪场收集,另外两株从另外两个无已知关联的猪场收集。
15种血清型参考菌株均给出了独特、可重复的ERIC-PCR指纹图谱。来自三个疑似相关暴发的12株分离株均给出了单一指纹图谱,该图谱与15种血清型参考菌株以及研究猪场中的另外两株澳大利亚田间分离株所观察到的任何指纹图谱均不同。确认所有12株分离株为同一菌株后,制定了预防和控制计划,该计划已防止这三个猪场再次出现格氏病暴发。
ERIC-PCR是区分副猪嗜血杆菌不相关菌株的合适技术。12株副猪嗜血杆菌田间分离株具有相同指纹图谱这一发现有力地证明了这三个猪场存在共同感染源。本研究首次表明,ERIC-PCR是副猪嗜血杆菌亚型分型的合适技术,对研究格氏病暴发的流行病学有用。