Serraino A, Marchetti G, Sanguinetti V, Rossi M C, Zanoni R G, Catozzi L, Bandera A, Dini W, Mignone W, Franzetti F, Gori A
Department of Veterinary Public Health and Animal Pathology, University of Bologna, Bologna, Italy.
J Clin Microbiol. 1999 Sep;37(9):2766-71. doi: 10.1128/JCM.37.9.2766-2771.1999.
An epidemiological survey for the monitoring of bovine tuberculosis transmission was carried out in western Liguria, a region in northern Italy. Fifteen Mycobacterium bovis strains were isolated from 63 wild boar samples (62 from mandibular lymph nodes and 1 from a liver specimen). Sixteen mediastinal lymph nodes of 16 head of cattle were collected, and 15 Mycobacterium bovis strains were subsequently cultured. All M. bovis strains isolated from cattle and wild boars were genotyped by spoligotyping and by restriction fragment length polymorphism (RFLP) analysis with the IS6110 and IS1081 probes. All M. bovis strains showed the typical spoligotype characterized by the absence of the 39 to 43 spacers in comparison with the number in M. tuberculosis. A total of nine different clusters were identified by spoligotyping. The largest cluster included 9 strains isolated from wild boars and 11 strains isolated from cattle, thus confirming the possibility of transmission between the two animal species. Fingerprinting by RFLP analysis with the IS6110 probe showed an identical single-band pattern for 29 of 30 strains analyzed, and only 1 strain presented a five-band pattern. The use of IS1081 as a second probe was useful for differentiation of M. bovis from M. bovis BCG but not for differentiation among M. bovis strains, which presented the same undifferentiated genomic profile. In relation to the epidemiological investigation, we hypothesized that the feeding in pastures contaminated by cattle discharges could represent the most probable route of transmission of M. bovis between the two animal species. In conclusion, our results confirmed the higher discriminatory power of spoligotyping in relation to that of RFLP analysis for the differentiation of M. bovis genomic profiles. Our data showed the presence of a common M. bovis genotype in both cattle and wild boars, confirming the possible interspecies transmission of M. bovis.
在意大利北部的利古里亚西部开展了一项用于监测牛结核病传播的流行病学调查。从63份野猪样本（62份来自下颌淋巴结，1份来自肝脏标本）中分离出15株牛分枝杆菌菌株。采集了16头牛的16个纵隔淋巴结，随后培养出15株牛分枝杆菌菌株。通过间隔寡核苷酸分型法（spoligotyping）以及使用IS6110和IS1081探针的限制性片段长度多态性（RFLP）分析，对从牛和野猪中分离出的所有牛分枝杆菌菌株进行基因分型。与结核分枝杆菌相比，所有牛分枝杆菌菌株均呈现典型的间隔寡核苷酸分型特征，即缺少39至43个间隔区。通过间隔寡核苷酸分型法共鉴定出9个不同的簇。最大的簇包括9株从野猪中分离出的菌株和11株从牛中分离出的菌株，从而证实了两种动物之间传播的可能性。使用IS6110探针进行RFLP分析的指纹图谱显示，在分析的30株菌株中有29株呈现相同的单带模式，只有1株呈现五带模式。使用IS1081作为第二种探针有助于区分牛分枝杆菌和卡介苗，但对于区分牛分枝杆菌菌株并无帮助，这些菌株呈现相同的未分化基因组图谱。关于流行病学调查，我们推测在被牛粪污染的牧场中觅食可能是牛分枝杆菌在两种动物之间传播的最可能途径。总之，我们的结果证实，与RFLP分析相比，间隔寡核苷酸分型法在区分牛分枝杆菌基因组图谱方面具有更高的鉴别力。我们的数据显示牛和野猪中均存在共同的牛分枝杆菌基因型，证实了牛分枝杆菌可能存在种间传播。
