Sardelić Sanda, Fournier Pierre-Edouard, Punda Polić Volga, Bradarić Nikola, Grgić Dusanka, Ivić Ivo, Ledina Dragan, Luksić Boris, Milas Ivo, Raoult Didier
Department of Clinical Microbiology, Split University Hospital, Croatia.
Croat Med J. 2003 Oct;44(5):630-4.
To detect and isolate rickettsial strains from blood samples of patients with presumptive diagnosis of Mediterranean spotted fever (MSF) in the coastal region of south Croatia, and to compare the results with routine serology.
A "suicide" polymerase chain reaction (PCR), and a shell vial culture were done on samples of ethylenediamine tetra-acetic acid (EDTA) and citrate-anticoagulated blood samples. Indirect immunofluorescence was performed on sera collected from 17 patients clinically diagnosed with MSF during summer in three consecutive years, from 1998 to 2000.
The primers used in PCR amplified the expected part of the rickettsia genomic DNA and Rickettsia conorii grew from the shell vial-cultured blood of a single patient. In 13 patients, the diagnosis was confirmed serologically by paired sera, whereas in 4 patients the diagnosis remained presumptive, since no paired sera were available. Analyzing sequences of the ompA and citrate synthase gene, respectively, derived from the shell vial isolate, a 100% similarity with Rickettsia conorii, strain Seven (Malish), was found.
To the best of our knowledge, this is the first isolation of Rickettsia conorii from a human sample in Croatia, and the first proof of a causative agent of MSF in the country. Beside PCR-based methods and isolation, correct diagnosis of MSF could be still routinely reached by serology.
从克罗地亚南部沿海地区疑似诊断为地中海斑疹热（MSF）患者的血样中检测和分离立克次体菌株，并将结果与常规血清学进行比较。
对乙二胺四乙酸（EDTA）和柠檬酸盐抗凝的血样进行“自杀”聚合酶链反应（PCR）和空斑培养。对1998年至2000年连续三年夏季临床诊断为MSF的17例患者的血清进行间接免疫荧光检测。
PCR中使用的引物扩增了立克次体基因组DNA的预期部分，并且从一名患者的空斑培养血中培养出了康氏立克次体。13例患者通过双份血清在血清学上得到确诊，而4例患者由于没有双份血清，诊断仍为疑似。分别分析来自空斑分离株的ompA和柠檬酸合酶基因序列，发现与康氏立克次体菌株七号（Malish）有100%的相似性。
据我们所知，这是克罗地亚首次从人类样本中分离出康氏立克次体，也是该国MSF病原体的首个证据。除了基于PCR的方法和分离外，通过血清学仍可常规实现MSF的正确诊断。
