Motiwala Alifiya S, Strother Megan, Amonsin Alongkorn, Byrum Beverly, Naser Saleh A, Stabel Judith R, Shulaw William P, Bannantine John P, Kapur Vivek, Sreevatsan Srinand
Food Animal Health Research Program, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, 44691, USA.
J Clin Microbiol. 2003 May;41(5):2015-26. doi: 10.1128/JCM.41.5.2015-2026.2003.
The objectives of this study were to understand the molecular diversity of animal and human strains of Mycobacterium avium subsp. paratuberculosis isolated in the United States and to identify M. avium subsp. paratuberculosis-specific diagnostic molecular markers to aid in disease detection, prevention, and control. Multiplex PCR of IS900 integration loci (MPIL) and amplified fragment length polymorphism (AFLP) analyses were used to fingerprint M. avium subsp. paratuberculosis isolates recovered from animals (n = 203) and patients with Crohn's disease (n = 7) from diverse geographic localities. Six hundred bacterial cultures, including M. avium subsp. paratuberculosis (n = 303), non-M. avium subsp. paratuberculosis mycobacteria (n = 129), and other nonmycobacterial species (n = 168), were analyzed to evaluate the specificity of two IS900 integration loci and a newly described M. avium subsp. paratuberculosis-specific sequence (locus 251) as potential targets for the diagnosis of M. avium subsp. paratuberculosis. MPIL fingerprint analysis revealed that 78% of bovine origin M. avium subsp. paratuberculosis isolates clustered together into a major node, whereas isolates from human and ovine sources showed greater genetic diversity. MPIL analysis also showed that the M. avium subsp. paratuberculosis isolates from ovine and bovine sources from the same state were more closely associated than were isolates from different geographic regions, suggesting that some of the strains are shared between these ruminant species. AFLP fingerprinting revealed a similar pattern, with most isolates from bovine sources clustering into two major nodes, while those recovered from sheep or humans were clustered on distinct branches. Overall, this study identified a high degree of genetic similarity between M. avium subsp. paratuberculosis strains recovered from cows regardless of geographic origin. Further, the results of our analyses reveal a relatively higher degree of genetic heterogeneity among M. avium subsp. paratuberculosis isolates recovered from human and ovine sources.
本研究的目的是了解在美国分离出的禽分枝杆菌副结核亚种动物和人类菌株的分子多样性，并鉴定禽分枝杆菌副结核亚种特异性诊断分子标记，以协助疾病的检测、预防和控制。采用IS900整合位点多重PCR（MPIL）和扩增片段长度多态性（AFLP）分析对从不同地理区域的动物（n = 203）和克罗恩病患者（n = 7）中分离出的禽分枝杆菌副结核亚种菌株进行指纹分析。对600株细菌培养物进行分析，包括禽分枝杆菌副结核亚种（n = 303）、非禽分枝杆菌副结核亚种分枝杆菌（n = 129）和其他非分枝杆菌物种（n = 168），以评估两个IS900整合位点和一个新描述的禽分枝杆菌副结核亚种特异性序列（位点251）作为禽分枝杆菌副结核亚种诊断潜在靶点的特异性。MPIL指纹分析显示，78%源自牛的禽分枝杆菌副结核亚种菌株聚集在一个主要节点，而源自人类和绵羊的菌株显示出更大的遗传多样性。MPIL分析还表明，来自同一州的绵羊和牛源的禽分枝杆菌副结核亚种菌株比来自不同地理区域的菌株关联更紧密，这表明这些反刍动物物种之间共享一些菌株。AFLP指纹分析显示出类似的模式，大多数源自牛的菌株聚集在两个主要节点，而从绵羊或人类中分离出的菌株则聚集在不同的分支上。总体而言，本研究发现，无论地理来源如何，从奶牛中分离出的禽分枝杆菌副结核亚种菌株之间存在高度的遗传相似性。此外，我们的分析结果显示，从人类和绵羊来源分离出的禽分枝杆菌副结核亚种菌株之间的遗传异质性相对较高。
