从墨西哥感染牛分枝杆菌的活牛鼻渗出液中直接制备DNA修饰的CTAB为人类结核病诊断试验提供了一种有价值的外推分析方法

Sanchez-Garza J.J, G. B.M., M. G.A, Cepeda Hortensia, Vázquez A.J, F. J.M., Guerrero Gg
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引用次数: 0

摘要

MTB复合体成员的检测、鉴定和分化依赖于自90年代以来发展起来的方法的特异性、敏感性和准确性。尽管如此,在发展中国家的流行地区,主要由于费用和可获得性的原因,仍然进行结核菌素现场试验以及常规技术(组织病理学和细菌学)。因此,迫切需要一种常规的检测方法,以提高活牛的现场检测(假阳性和阴性检测),同时避免不必要的动物牺牲。为此,在目前的工作中,我们设计了一种双重实验策略,可以作为常规检测方法,通过PCR介导的RD 's扩增检测牛分枝杆菌或结核分枝杆菌。DNA可以从快速生长的菌落(7 - 8天)或匀浆组织、鼻渗出液和纯化介导的十六烷基三甲基溴化铵(CTAB)阳离子缓冲液中制备。该方法适用于结核阳性人鼻/口渗出液。结果相似。总的来说,这些发现表明,这一战略是TBb流行病学调查和研究的一个有价值的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Direct DNA Modified CTAB Preparation from Nasal Exudate in Live M. bovis Infected Cattle in Mexico Provide with a Valuable Assay Extrapolated to Humans TB Diagnostic Test
Detection, identification, and differentiation of members of the MTB complex rely on specificity, sensitivity, and accuracy of the methods that have been developed since the decades of the ’90s. Despite this, still, in endemic areas of developing countries tuberculin field test as well as conventional techniques (histopathology and bacteriology) are performed due primarily to the costs and availability. Therefore, it is an urgent need to have a routine assay to boost field test (false positive and negative tests) in live cows while avoiding the unnecessary sacrifice of animals. To this end, in the present work, we designed a dual experimental strategy that can be used as a routine assay for the M. bovis or M. tuberculosis detection through PCR mediated amplification of RD’s. DNA can be prepared from fast-growing colonies (7 to 8 days) or from homogenized tissue, nasal exudate and purification mediated cetyl-trimethylammonium bromide (CTAB) cationic buffer. The method was extraplated to positive TB positive nasal/oral human exudate.with similar results. Collectively these findings indicate that this strategy represent a valuable tool for TBb epidemiological survey and research.
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