Lee Sin Hang, Vigliotti Jessica S, Vigliotti Veronica S, Jones William, Moorcroft Thomas A, Lantsman Katherine
Department of Pathology, Milford Hospital, 300 Seaside Ave., Milford, CT 06460, USA.
Origins of Health, LLC, 279 New Britain Road Berlin, CT 06037, USA.
Int J Mol Sci. 2014 Jun 25;15(7):11364-86. doi: 10.3390/ijms150711364.
A highly conserved 357-bp segment of the 16S ribosomal RNA gene (16S rDNA) of Borrelia burgdorferi sensu lato and the correspondent 358-bp segment of the Borrelia miyamotoi gene were amplified by a single pair of nested polymerase chain reaction (PCR) primers for detection, and the amplicons were used as the templates for direct Sanger DNA sequencing. Reliable molecular diagnosis of these borreliae was confirmed by sequence alignment analysis of the hypervariable regions of the PCR amplicon, using the Basic Local Alignment Search Tool (BLAST) provided by the GenBank. This methodology can detect and confirm B. burgdorferi and B. miyamotoi in blood samples of patients with off-season spirochetemia of low bacterial density. We found four B. miyamotoi infections among 14 patients with spirochetemia, including one patient co-infected by both B. miyamotoi and B. burgdorferi in a winter month when human exposure to tick bites is very limited in the Northeast of the U.S.A. We conclude that sensitive and reliable tests for these two Borrelia species should be implemented in the microbiology laboratory of hospitals located in the disease-endemic areas, for timely diagnosis and appropriate treatment of the patients at an early stage of the infection to prevent potential tissue damages.
使用一对巢式聚合酶链反应（PCR）引物扩增伯氏疏螺旋体狭义种16S核糖体RNA基因（16S rDNA）高度保守的357 bp片段以及宫本疏螺旋体基因对应的358 bp片段用于检测，扩增产物用作直接桑格DNA测序的模板。通过使用GenBank提供的基本局部比对搜索工具（BLAST）对PCR扩增产物高变区进行序列比对分析，证实了对这些疏螺旋体的可靠分子诊断。该方法可检测并确认低细菌密度的非季节性螺旋体血症患者血样中的伯氏疏螺旋体和宫本疏螺旋体。我们在14例螺旋体血症患者中发现了4例宫本疏螺旋体感染，其中1例患者在冬季同时感染了宫本疏螺旋体和伯氏疏螺旋体，当时在美国东北部人类接触蜱叮咬的情况非常有限。我们得出结论，在疾病流行地区的医院微生物实验室应开展针对这两种疏螺旋体的灵敏且可靠的检测，以便在感染早期及时诊断并适当治疗患者，防止潜在的组织损伤。
