Zakharova Irina, Teteryatnikova Natalya, Toporkov Andrey, Viktorov Dmitry
Department of Microbiology, Volgograd Plague Control Research Institute, Volgograd, 400131, Russian Federation.
Department of Microbiology, Volgograd Plague Control Research Institute, Volgograd, 400131, Russian Federation.
Acta Trop. 2017 Oct;174:1-8. doi: 10.1016/j.actatropica.2017.06.016. Epub 2017 Jun 17.
Two species of Burkholderia pseudomallei complex (Bpc), B. pseudomallei and B. mallei, can cause severe life-threatening infections. Rapidly discerning individual species within the group and separating them from other opportunistic pathogens of the Burkholderia cepacia complex (Bcc) is essential to establish a correct diagnosis and for epidemiological surveillance. In this study, a multiplex PCR assay based on the detection of an individual set of chromosomal beta-lactamase genes for single-step identification and differentiation of B. pseudomallei, B. mallei, B. thailandensis, and Bcc was developed. Two pairs of primers specific to a distinct class of B metallo-beta-lactamase genes and a pair of primers specific to the oxacillin-hydrolyzing class D beta-lactamase gene were demonstrated to successfully discriminate species within Bpc and from Bcc. The assay sensitivity was 9561 genomic equivalents (GE) for B. pseudomallei, 7827 GE for B. mallei, 8749 GE for B. thailandensis and 6023 GE for B. cepacia.
两种类鼻疽伯克霍尔德菌复合体（Bpc），即类鼻疽伯克霍尔德菌和鼻疽伯克霍尔德菌，可引起严重的危及生命的感染。快速鉴别该组内的各个菌种并将它们与洋葱伯克霍尔德菌复合体（Bcc）的其他机会致病菌区分开来，对于确立正确诊断和进行流行病学监测至关重要。在本研究中，开发了一种基于检测一组单个染色体β-内酰胺酶基因的多重PCR检测方法，用于一步法鉴定和区分类鼻疽伯克霍尔德菌、鼻疽伯克霍尔德菌、泰国伯克霍尔德菌和Bcc。两对特异性针对不同类别的B型金属β-内酰胺酶基因的引物和一对特异性针对水解苯唑西林的D类β-内酰胺酶基因的引物被证明能够成功区分Bpc内的菌种以及与Bcc的菌种。该检测方法的灵敏度对于类鼻疽伯克霍尔德菌为9561个基因组当量（GE），对于鼻疽伯克霍尔德菌为7827 GE，对于泰国伯克霍尔德菌为8749 GE，对于洋葱伯克霍尔德菌为6023 GE。
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