Molecular detection and analysis of spotted fever group Rickettsia in patients with fever and rash at a tertiary care centre in Tamil Nadu, India
Molecular detection and analysis of spotted fever group Rickettsia in patients with fever and rash at a tertiary care centre in Tamil Nadu, India
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DOI：
10.1179/2047773212y.0000000001
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发表时间：
2012-03-01
影响因子：
3.4
通讯作者：
Dumler, John Stephen
中科院分区：
文献类型：
--
作者：
Prakash, John Antony Jude;Lal, T. Sohan;Dumler, John Stephen
背景：用聚合酶链式反应检测特定靶点来确认斑点热的诊断，但血清学试验仍被广泛使用。方法：对58例临床疑似斑点热患者，用套式聚合酶链式反应检测皮疹组织中GLTA、17 kDa脂蛋白抗原基因(17 KDa)、ompA和ompB。对代表四个靶点的扩增产物进行测序，以确认扩增的特异性。结果：从38例、23例、27例和22例皮肤活检标本中分别检测到GLTA、17 kDa、ompA和ompB基因。序列分析表明，GLTA、17 kDa、OmpA和ompB序列属于斑点热立克次体。在6个OmpA基因的部分序列中，只有1个与以前报道的‘Candidatus Rickettsia kellyi’不同。结论：进一步的证据表明，与‘Candidatus Rickettsia kellyi’相似的SFG立克次体在印度南部引起发热和皮疹。在分离培养中的立克次体后，需要进行更详细的系统发育分析，以便为该地区发生新型斑点热立克次体提供确凿的证据。
Background: Detection of specific targets by PCR is used to confirm a diagnosis of spotted fever, but serological tests are still widely used. In this prospective study, nested PCR was performed on skin biopsy specimens to confirm the diagnosis of spotted fever.Methods: In 58 clinically suspected cases of spotted fever, nested PCR, to detect gltA, 17 kDa lipoprotein antigen gene (17 kDa), ompA and ompB, from skin biopsy of the rash was performed. Sequencing was carried on amplicons representing the four targets to confirm specificity of amplification. This was followed by phylogenetic analysis using MEGA version 4.0 software.Results: The gltA, 17 kDa, ompA, and ompB genes were detected from skin biopsy specimens in 38, 23, 27, and 22 individuals. Sequence analysis revealed that the gltA, 17 kDa, ompA, and ompB sequences belonged to spotted fever group (SFG) rickettsia. Of the six partial ompA gene sequences, only one was dissimilar to the previously reported 'Candidatus Rickettsia kellyi'.Conclusion: Further evidence indicates that SFG rickettsiae resembling 'Candidatus Rickettsia kellyi' cause fever and rash in southern India. More detailed phylogenetic analysis following isolation of rickettsia in culture is required for providing irrefutable proof for the occurrence of novel spotted fever rickettsiae in this region.