Leth Christoph, Varadharajan Ashok, Mester Patrick, Fischaleck Marlis, Rossmanith Peter, Schmoll Friedrich, Fink Maria
Austrian Agency for Health and Food Safety, Institute for Veterinary Disease Control, Moedling, Lower Austria, Austria.
University of Applied Sciences FH Campus Wien, Vienna, Austria.
PLoS One. 2017 Jul 19;12(7):e0181157. doi: 10.1371/journal.pone.0181157. eCollection 2017.
Mycobacterium caprae, a member of the Mycobacterium tuberculosis complex, is the main causative agent of bovine tuberculosis in alpine regions. Bacterial culture is the gold standard in bovine tuberculosis diagnostic but takes up to twelve weeks. This increases the time and costs for stocks affected with bovine tuberculosis. Hence this study focused on the implementation of a fast and precise mycobacterial detection method and compared it with currently used methods. Matrix lysis is a chemical lysis using high concentrations of urea to solubilize bovine and red deer tissue and was used to detect even smallest amounts or non-visible lesions of mycobacteria. A total of 64 samples collected from 44 animals (37 red deer and 7 cattle) were tested by Matrix lysis. Forty-three of these samples were used for Mycobacterium tuberculosis complex detection by quantitative PCR and other 21 for subtyping the genetically different variants of M. caprae. Furthermore, three Matrix lysis samples were used for Next Generation Sequencing. Our results confirm that Matrix lysis is a fast and precise method for detecting Mycobacterium tuberculosis complex in native tissue samples. However, at the moment it reaches its limits when the samples were analyzed by Next Generation Sequencing and RD4 subtyping.
山羊分枝杆菌是结核分枝杆菌复合群的成员之一，是高寒地区牛结核病的主要病原体。细菌培养是牛结核病诊断的金标准，但需要长达12周的时间。这增加了受牛结核病影响的畜群的时间和成本。因此，本研究重点关注一种快速、精确的分枝杆菌检测方法的实施，并将其与目前使用的方法进行比较。基质裂解是一种使用高浓度尿素溶解牛和马鹿组织的化学裂解方法，用于检测分枝杆菌的最小量或不可见病变。通过基质裂解对从44只动物（37只马鹿和7头牛）采集的64个样本进行了检测。其中43个样本用于通过定量PCR检测结核分枝杆菌复合群，另外21个样本用于对山羊分枝杆菌的基因不同变体进行亚型分析。此外，3个基质裂解样本用于下一代测序。我们的结果证实，基质裂解是一种在天然组织样本中检测结核分枝杆菌复合群的快速、精确方法。然而，目前当通过下一代测序和RD4亚型分析对样本进行分析时，它达到了其局限性。
