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波斯疏螺旋体:通过常规PCR以及对从以色列一只猫和蜱虫中分离出的两株菌株进行多位点序列分析实现体外培养及特性鉴定

Borrelia persica: In vitro cultivation and characterization via conventional PCR and multilocus sequence analysis of two strains isolated from a cat and ticks from Israel.

作者信息

Schwarzer Sandra, Margos Gabriele, Overzier Evelyn, Fingerle Volker, Baneth Gad, Straubinger Reinhard K

机构信息

Bacteriology and Mycology, Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-Universität München, Veterinärstraße 13, 80539 München, Germany.

German National Reference Centre for Borrelia, Bavarian Health and Food Safety Authority, Veterinärstraße 2, 85764 Oberschleißheim, Germany.

出版信息

Ticks Tick Borne Dis. 2015 Sep;6(6):751-7. doi: 10.1016/j.ttbdis.2015.06.012. Epub 2015 Jul 3.

Abstract

Borrelia persica, one of the pathogenic agents of tick-borne relapsing fever, is transmitted by the soft tick Ornithodoros tholozani. It causes infections in humans as well as in animals. In this study, we developed a medium, termed Pettenkofer/LMU Bp, for reliable in vitro cultivation. Cell densities up to 5.2×10(7) viable cells/ml were achieved over at least 40 passages. The cultivable B. persica strain isolated from a cat was further analyzed by amplification of the flaB gene using conventional PCR. In addition, seven housekeeping genes (clpA, clpX, pepX, pyrG, recG, rplB and uvrA) of this B. persica strain and a second strain isolated out of pooled ticks from Israel were amplified and the phylogenetic relationships among Borrelia species were analyzed. The results of the conventional PCR and the multilocus sequence analysis confirmed our isolates as B. persica.

摘要

波斯疏螺旋体是蜱传回归热的病原体之一,由软蜱托氏钝缘蜱传播。它可导致人类和动物感染。在本研究中,我们开发了一种名为Pettenkofer/LMU Bp的培养基,用于可靠的体外培养。在至少40代培养过程中,细胞密度达到了5.2×10⁷个活细胞/毫升。使用常规PCR扩增flaB基因,对从一只猫身上分离出的可培养波斯疏螺旋体菌株进行了进一步分析。此外,对该波斯疏螺旋体菌株以及从以色列采集的混合蜱中分离出的另一菌株的七个管家基因(clpA、clpX、pepX、pyrG、recG、rplB和uvrA)进行了扩增,并分析了疏螺旋体物种之间的系统发育关系。常规PCR和多位点序列分析结果证实我们分离出的菌株为波斯疏螺旋体。

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