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利用随机扩增多态性DNA-PCR和23S rRNA-PCR对肉鸡群空肠弯曲菌进行分子流行病学研究及垫料在其传播中的作用

Molecular epidemiology of Campylobacter jejuni in broiler flocks using randomly amplified polymorphic DNA-PCR and 23S rRNA-PCR and role of litter in its transmission.

作者信息

Payne R E, Lee M D, Dreesen D W, Barnhart H M

机构信息

Department of Environmental Health, College of Veterinary Medicine, University of Georgia, Athens, Georgia 30602-2102, USA.

出版信息

Appl Environ Microbiol. 1999 Jan;65(1):260-3. doi: 10.1128/AEM.65.1.260-263.1999.

Abstract

Poultry has long been cited as a reservoir for Campylobacter spp., and litter has been implicated as a vehicle in their transmission. Chicks were raised on litter removed from a broiler house positive for Campylobacter jejuni. Litter was removed from the house on days 0, 3, and 9 after birds were removed for slaughter. Chicks were raised on these three litters under controlled conditions in flocks of 25. None of these birds yielded C. jejuni in their cecal droppings through 7 weeks. Two successive flocks from the same Campylobacter-positive broiler house were monitored for Campylobacter colonization. Campylobacter jejuni prevalence rates were determined for each flock. Randomly amplified polymorphic DNA (RAPD)-PCR and 23S rRNA-PCR typing methods were used to group isolates. A high prevalence (60%) of C. jejuni in flock 1 coincided with the presence of an RAPD profile not appearing in flock 2, which had a lower rate of prevalence (28%). A 23S rRNA-PCR typing method was used to determine if strains with different RAPD profiles and different prevalence rates contained different 23S sequences. RAPD profiles detected with higher prevalence rates contained a spacer in the 23S rRNA region 100% of the time, while RAPD profiles found with lower prevalence rates contained an intervening sequence less than 2% of the time. Data suggest varying colonizing potentials of different RAPD profiles and a source other than previously used litter as a means of transmission of C. jejuni. These molecular typing methods demonstrate their usefulness, when used together, in this epidemiologic investigation.

摘要

长期以来,家禽一直被认为是弯曲杆菌属的宿主,垫料被认为是其传播的媒介。雏鸡在从空肠弯曲菌呈阳性的肉鸡舍中取出的垫料上饲养。在鸡只被送去屠宰后的第0、3和9天,从鸡舍中取出垫料。雏鸡在这三种垫料上,在可控条件下,以每群25只的规模饲养。在7周内,这些鸡的盲肠粪便中均未检出空肠弯曲菌。对来自同一空肠弯曲菌阳性肉鸡舍的连续两批鸡群进行了空肠弯曲菌定植监测。测定了每批鸡群的空肠弯曲菌流行率。使用随机扩增多态性DNA(RAPD)-PCR和23S rRNA-PCR分型方法对分离株进行分组。第1批鸡群中空肠弯曲菌的高流行率(60%)与第2批鸡群中未出现的RAPD图谱一致,第2批鸡群的流行率较低(28%)。使用23S rRNA-PCR分型方法来确定具有不同RAPD图谱和不同流行率的菌株是否包含不同的23S序列。流行率较高的RAPD图谱在23S rRNA区域100%的时间内含有一个间隔区,而流行率较低的RAPD图谱在不到2%的时间内含有一个间隔序列。数据表明不同RAPD图谱的定植潜力不同,并且空肠弯曲菌的传播途径除了先前使用的垫料外还有其他来源。这些分子分型方法在这项流行病学调查中一起使用时证明了它们的实用性。

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