Beinhauerova Monika, Slana Iva
Department of Microbiology and Antimicrobial Resistance, Veterinary Research Institute, Hudcova 70, 62100 Brno, Czech Republic.
Department of Experimental Biology, Faculty of Science, Masaryk University, Kamenice 5, 62500 Brno, Czech Republic.
Microorganisms. 2021 Jan 23;9(2):237. doi: 10.3390/microorganisms9020237.
An important prerequisite for the effective control, timely diagnosis, and successful treatment of mycobacterial infections in both humans and animals is a rapid, specific, and sensitive detection technique. Culture is still considered the gold standard in the detection of viable mycobacteria; however, mycobacteria are extremely fastidious and slow-growing microorganisms, and therefore cultivation requires a very long incubation period to obtain results. Polymerase Chain Reaction (PCR) methods are also frequently used in the diagnosis of mycobacterial infections, providing faster and more accurate results, but are unable to distinguish between a viable and non-viable microorganism, which results in an inability to determine the success of tuberculosis patient treatment or to differentiate between an active and passive infection of animals. One suitable technique that overcomes these shortcomings mentioned is the phage amplification assay (PA). PA specifically detects viable mycobacteria present in a sample within 48 h using a lytic bacteriophage isolated from the environment. Nowadays, an alternative approach to PA, a commercial kit called Actiphage™, is also employed, providing the result within 6-8 h. In this approach, the bacteriophage is used to lyse mycobacterial cells present in the sample, and the released DNA is subsequently detected by PCR. The objective of this review is to summarize information based on the PA used for detection of mycobacteria significant in both human and veterinary medicine from various kinds of matrices.
对人类和动物的分枝杆菌感染进行有效控制、及时诊断和成功治疗的一个重要前提是要有快速、特异且灵敏的检测技术。培养法仍是检测活分枝杆菌的金标准；然而，分枝杆菌是极其苛求且生长缓慢的微生物，因此培养需要很长的孵育期才能获得结果。聚合酶链反应（PCR）方法也常用于分枝杆菌感染的诊断，能提供更快且更准确的结果，但无法区分活的和非活的微生物，这导致无法确定结核病患者的治疗效果，也无法区分动物的主动感染和被动感染。一种克服上述这些缺点的合适技术是噬菌体扩增试验（PA）。PA使用从环境中分离出的裂解性噬菌体在48小时内特异性检测样品中存在的活分枝杆菌。如今，PA的一种替代方法，即一种名为Actiphage™的商业试剂盒也被采用，它能在6 - 8小时内给出结果。在这种方法中，噬菌体用于裂解样品中存在的分枝杆菌细胞，随后通过PCR检测释放的DNA。本综述的目的是基于用于从各种基质中检测在人类和兽医学中都很重要的分枝杆菌的PA来总结信息。
