Thoresen O F, Olsaker I
National Veterinary Institute, Oslo, Norway.
Vet Microbiol. 1994 Jun;40(3-4):293-303. doi: 10.1016/0378-1135(94)90118-x.
Reference strains and 31 clinical isolates of M. paratuberculosis, mainly from goats, were analysed for restriction fragment length polymorphism (RFLP). Restriction digests of bacterial DNA were hybridized with a repetitive insertion sequence, IS900, to obtain banding patterns for comparison of strains. Twenty-five of the 31 field-strains hybridized with IS900, and five hybridization patterns were identified. It was not possible to identify specific patterns for goat strains of M. paratuberculosis. Four hybridization patterns were similar, whereas the fifth pattern of a sheep strain diverged considerably in position and number of bands. Six goat strains failed to hybridize with IS900, and the absence of IS900 was verified by the polymerase chain reaction and hybridization with an oligonucleotide probe. The six IS900-negative goat strains had diverging phenotypic properties, and the identification of these strains is discussed. The present study shows that M. paratuberculosis strains infecting goats are genetically similar to cattle strains and that IS900 is a specific genetic element for identification of M. paratuberculosis.
对副结核分枝杆菌的参考菌株以及31株临床分离株（主要来自山羊）进行了限制性片段长度多态性（RFLP）分析。细菌DNA的限制性酶切产物与重复插入序列IS900杂交，以获得菌株比较的条带模式。31株野外菌株中的25株与IS900杂交，并鉴定出5种杂交模式。无法鉴定出副结核分枝杆菌山羊菌株的特定模式。4种杂交模式相似，而绵羊菌株的第5种模式在条带位置和数量上有很大差异。6株山羊菌株未与IS900杂交，通过聚合酶链反应和与寡核苷酸探针杂交验证了IS900的缺失。这6株IS900阴性山羊菌株具有不同的表型特性，并对这些菌株的鉴定进行了讨论。本研究表明，感染山羊的副结核分枝杆菌菌株在基因上与牛菌株相似，且IS900是鉴定副结核分枝杆菌的一种特异性遗传元件。
