Caimi K, Romano M I, Alito A, Zumarraga M, Bigi F, Cataldi A
Instituto de Biotecnología, Centro de Investigaciones en Ciencias Veterinarias, Instituto Nacional de Tecnología Agropecuaria, 1708 Moron, Argentina.
J Clin Microbiol. 2001 Mar;39(3):1067-72. doi: 10.1128/JCM.39.3.1067-1072.2001.
Spoligotyping is a major tool for molecular typing of Mycobacterium bovis. This technique is based on the polymorphism of spacers that separate direct repeats (DRs) in the M. tuberculosis complex DR region. Numerous M. bovis strains show a lack of several spacers which appears as a gap in the spoligotyping pattern. To determine whether these gaps contain alternative spacers not included in the spoligotyping membrane, PCRs using primers that hybridize to the spacers adjacent to the gaps were performed. Comparing the sizes of products obtained by PCR with those deduced from spoligotyping patterns, fragments were selected and sequenced to look for alternative spacers. Upon analysis of the sequences, five alternative spacers were detected, although deletions of spacers are mainly responsible for the observed gaps. The alternative spacers, which are more frequent in M. bovis than in M. tuberculosis, may contribute to increased M. bovis differentiation.
间隔寡核苷酸分型是牛分枝杆菌分子分型的主要工具。该技术基于结核分枝杆菌复合群直接重复序列（DR）区域中分隔直接重复序列的间隔序列的多态性。许多牛分枝杆菌菌株显示出几个间隔序列缺失，这在间隔寡核苷酸分型图谱中表现为一个缺口。为了确定这些缺口中是否包含间隔寡核苷酸分型膜上未包含的替代间隔序列，使用与缺口相邻的间隔序列杂交的引物进行了聚合酶链反应（PCR）。将PCR获得的产物大小与间隔寡核苷酸分型图谱推导的大小进行比较，选择片段并测序以寻找替代间隔序列。通过对序列的分析，检测到五个替代间隔序列，尽管间隔序列的缺失是观察到的缺口的主要原因。这些替代间隔序列在牛分枝杆菌中比在结核分枝杆菌中更常见，可能有助于增加牛分枝杆菌的鉴别力。
