Hulínská D, Votýpka J, Vanousová D, Hercogová J, Hulínský V, Drevová H, Kurzová Z, Uherková L
National Reference Laboratory, WHO Centre for Reference and Research on Borreliosis, National Institute of Public Health, Prague, Czech Republic.
Folia Microbiol (Praha). 2009;54(3):246-56. doi: 10.1007/s12223-009-0039-0. Epub 2009 Aug 2.
Anaplasma phagocytophilum has been first isolated from the blood of two Czech patients simultaneously with a cultivation of Borrelia burgdorferi sensu lato from their erythema migrans lesions. Cultivation of different Borrelia spp. from 12 erythema migrans biopsies, from 2 blood, one liquor and one placenta sample in BSK-H medium was successful. Adapted conventional methods targeting 16S rRNA and OspA genes for real-time polymerase chain reaction (PCR) and partial sequencing of these genes together with microscopical examinations of the blood smears provided a direct detection of the B. afzelii, B. burgdorferi, B. garinii, B. valaisiana and B. bissettii in the skin, B. garinii in the blood, placenta and liquor in 24 (36.3 %) patients, and A. phagocytophilum in 10 (15 %) patients with erythema migrans. Positive indirect IgM immunofluorescence against Anaplasma sp. was obtained in 7 cases, specific IgG antibodies were detected in 12 patients. Three women suffering from erythema migrans in the first trimester had positive PCR for Anaplasma and/or for Borrelia in the blood and two of them, later, in the placenta. Interpretation of laboratory data can bring important contribution to establishing the role of Anaplasma sp. in erythema migrans and forming the principle of precaution with laboratory diagnosis during pregnancy which always should be reflected in the resistance of Anaplasma sp. toward penicillins.
嗜吞噬细胞无形体最初是从两名捷克患者的血液中分离出来的，同时从他们的游走性红斑皮损中培养出了伯氏疏螺旋体狭义种。在BSK - H培养基中，从12份游走性红斑活检组织、2份血液、1份脑脊液和1份胎盘样本中成功培养出了不同的疏螺旋体物种。采用针对16S rRNA和OspA基因的改良常规方法进行实时聚合酶链反应（PCR）以及这些基因的部分测序，并结合血液涂片的显微镜检查，直接检测出24例（36.3%）患者的皮肤中有阿氏疏螺旋体、伯氏疏螺旋体、伽氏疏螺旋体、瓦氏疏螺旋体和比氏疏螺旋体，2例患者的血液、胎盘和脑脊液中有伽氏疏螺旋体，10例（15%）游走性红斑患者中有嗜吞噬细胞无形体。7例患者针对无形体属的间接IgM免疫荧光检测呈阳性，12例患者检测到特异性IgG抗体。3名在孕早期患有游走性红斑的女性血液中嗜吞噬细胞无形体和/或伯氏疏螺旋体的PCR检测呈阳性，其中2名女性后来在胎盘中检测到阳性。实验室数据的解读可为确定无形体属在游走性红斑中的作用以及形成孕期实验室诊断的预防原则提供重要贡献，这一原则应始终体现在无形体属对青霉素的耐药性上。
