Shen Z, Feng Y, Dewhirst F E, Fox J G
Division of Comparative Medicine, Massachusetts Institute of Technology, 77 Massachusetts Ave., Cambridge, MA 02139, USA.
J Clin Microbiol. 2001 Jun;39(6):2166-72. doi: 10.1128/JCM.39.6.2166-2172.2001.
During a 6-year period, 64 of 227 commercially reared cats had microaerobic bacteria isolated from their feces. All the isolates were initially identified as Campylobacter-like organisms based on biochemical and phenotypic characteristics. DNA extractions from 51 of these isolates were subjected to PCR using primers specific for Helicobacter spp. and Campylobacter spp. Of the isolates, 92% (47 of 51 isolates) were positive for Campylobacter spp., 41% (21 of 51 isolates) were positive for Helicobacter spp., 33% (17 of 51 isolates) were positive for both genera, 59% (30 of 51 isolates) were positive only for Campylobacter spp., and 8% (4 of 51) were positive only for Helicobacter spp. Sixteen of the 47 Campylobacter-positive cultures were positive for more than one Campylobacter spp. Based on a species-specific PCR assay, 83% of the isolates were identified as Campylobacter helveticus, 47% of the isolates were identified as Campylobacter upsaliensis, and 6% of the isolates were classified as Campylobacter jejuni. The 1.2-kb PCR products of the 16S rRNA genes of 19 Helicobacter species isolates were subjected to restriction fragment length polymorphism (RFLP) analysis. Of the five different RFLP patterns obtained, two clustered with Helicobacter ("Flexispira") taxon 8, one clustered with Helicobacter bilis, one clustered with Helicobacter canis, and the remaining pattern was closely related to a novel Helicobacter sp. strain isolated from a woodchuck. The sequence data for the 16S rRNA genes of 10 Helicobacter spp. validated the RFLP-based identification of these isolates. This study demonstrated that biochemical and phenotypic characteristics of microaerobic organisms in cat feces were insufficient to characterize mixed Helicobacter and Campylobacter infections. Molecular structure-based diagnostics using genus- and species-specific PCR, RFLP analysis, and 16S rRNA sequence analysis enabled the identification of multiple microaerobic species in individual animals. The clinical relevance of enteric Helicobacter and Campylobacter coinfection in cats will require further studies.
在6年期间,227只商业饲养的猫中有64只的粪便中分离出微需氧菌。根据生化和表型特征,所有分离株最初都被鉴定为弯曲菌样生物体。从其中51株分离株中提取DNA,使用针对幽门螺杆菌属和弯曲菌属的特异性引物进行PCR。在这些分离株中,92%(51株中的47株)弯曲菌属呈阳性,41%(51株中的21株)幽门螺杆菌属呈阳性,33%(51株中的17株)两个属均呈阳性,59%(51株中的30株)仅弯曲菌属呈阳性,8%(51株中的4株)仅幽门螺杆菌属呈阳性。47株弯曲菌阳性培养物中有16株对一种以上弯曲菌属呈阳性。基于种特异性PCR检测,83%的分离株被鉴定为瑞士弯曲菌,47%的分离株被鉴定为乌普萨拉弯曲菌,6%的分离株被分类为空肠弯曲菌。对19株幽门螺杆菌属分离株的16S rRNA基因的1.2kb PCR产物进行限制性片段长度多态性(RFLP)分析。在获得的五种不同RFLP模式中,两种与幽门螺杆菌(“Flexispira”)分类群8聚类,一种与胆汁幽门螺杆菌聚类,一种与犬幽门螺杆菌聚类,其余模式与从土拨鼠分离的一种新型幽门螺杆菌菌株密切相关。10株幽门螺杆菌属的16S rRNA基因的序列数据验证了基于RFLP对这些分离株的鉴定。这项研究表明,猫粪便中微需氧菌的生化和表型特征不足以表征幽门螺杆菌和弯曲菌的混合感染。使用属特异性和种特异性PCR、RFLP分析和16S rRNA序列分析的基于分子结构的诊断能够鉴定个体动物中的多种微需氧菌。猫肠道幽门螺杆菌和弯曲菌共感染的临床相关性需要进一步研究。