Knudsen K N, Bang D D, Nielsen E M, Madsen M
Department of Poultry, Fish and Fur Animals, Danish Institute for Food and Veterinary Research, Aarhus N, Denmark.
J Appl Microbiol. 2005;99(2):392-9. doi: 10.1111/j.1365-2672.2005.02594.x.
To apply and evaluate LG (LPS genes) genotyping, which is a genotyping method based on a cluster of genes involved in the synthesis of surface lipopolysaccharides (LPS) in Campylobacter species, for typing of Campylobacter jejuni isolates obtained from Danish broiler chickens. Furthermore, the LG genotyping method was used to study the genetic stability of four C. jejuni strains after gastrointestinal passage through experimentally infected chickens.
In the present study, the LG genotyping method was modified with respect to the restriction enzymes used. To validate the method, 63 Penner serotype reference strains and 107 C. jejuni chicken isolates, representing the most common Penner serotypes of C. jejuni in Danish poultry, were selected for typing. The method was successfully used for typing all isolates and the LG genotype profiles were reproducible. There were no changes in the LG genotype of the C. jejuni strains obtained after experimental passage through chickens.
All C. jejuni strains obtained from broiler chickens were typeable by the LG genotyping method. Application of the RsaI restriction enzyme improved the method in terms of ease and consistency of analyses and increase of discriminatory power.
The LG genotyping method is a valuable tool for typing C. jejuni isolates obtained from poultry. However, the association between Penner serotyping based on passive haemagglutination of heat-stable antigens and LG genotyping was low when applied to poultry isolates. This is in contrast to previous studies on isolates of human origin that reported a high correlation between results obtained by the two typing methods (Shi et al. 2002).
应用并评估LG(脂多糖基因)基因分型法,这是一种基于空肠弯曲菌属表面脂多糖(LPS)合成相关基因簇的基因分型方法,用于对从丹麦肉鸡中分离得到的空肠弯曲菌菌株进行分型。此外,LG基因分型法还用于研究四种空肠弯曲菌菌株经实验性感染鸡的胃肠道传代后的遗传稳定性。
在本研究中,对LG基因分型法所使用的限制性内切酶进行了改良。为验证该方法,选择了63株Penner血清型参考菌株和107株空肠弯曲菌鸡分离株进行分型,这些分离株代表了丹麦家禽中空肠弯曲菌最常见的Penner血清型。该方法成功用于所有分离株的分型,且LG基因型谱具有可重复性。经鸡实验传代后获得的空肠弯曲菌菌株的LG基因型没有变化。
从肉鸡中获得的所有空肠弯曲菌菌株都可用LG基因分型法进行分型。RsaI限制性内切酶的应用在分析的简便性和一致性以及鉴别力的提高方面改进了该方法。
LG基因分型法是对从家禽中分离得到的空肠弯曲菌菌株进行分型的有价值工具。然而,当应用于家禽分离株时,基于热稳定抗原被动血凝的Penner血清分型与LG基因分型之间的关联性较低。这与先前关于人类来源分离株的研究相反,后者报道这两种分型方法所得结果之间具有高度相关性(Shi等人,2002年)。