Lee Sin Hang, Vigliotti Veronica S, Vigliotti Jessica S, Jones William, Williams Jessie, Walshon Jay
Department of Pathology, Milford Hospital, 300 Seaside Avenue, Milford, 06460, USA.
BMC Res Notes. 2010 Nov 1;3:273. doi: 10.1186/1756-0500-3-273.
A sensitive and analytically specific nucleic acid amplification test (NAAT) is valuable in confirming the diagnosis of early Lyme disease at the stage of spirochetemia.
Venous blood drawn from patients with clinical presentations of Lyme disease was tested for the standard 2-tier screen and Western Blot serology assay for Lyme disease, and also by a nested polymerase chain reaction (PCR) for B. burgdorferi sensu lato 16S ribosomal DNA. The PCR amplicon was sequenced for B. burgdorferi genomic DNA validation. A total of 130 patients visiting emergency room (ER) or Walk-in clinic (WALKIN), and 333 patients referred through the private physicians' offices were studied. While 5.4% of the ER/WALKIN patients showed DNA evidence of spirochetemia, none (0%) of the patients referred from private physicians' offices were DNA-positive. In contrast, while 8.4% of the patients referred from private physicians' offices were positive for the 2-tier Lyme serology assay, only 1.5% of the ER/WALKIN patients were positive for this antibody test. The 2-tier serology assay missed 85.7% of the cases of early Lyme disease with spirochetemia. The latter diagnosis was confirmed by DNA sequencing.
Nested PCR followed by automated DNA sequencing is a valuable supplement to the standard 2-tier antibody assay in the diagnosis of early Lyme disease with spirochetemia. The best time to test for Lyme spirochetemia is when the patients living in the Lyme disease endemic areas develop unexplained symptoms or clinical manifestations that are consistent with Lyme disease early in the course of their illness.
一种灵敏且分析特异性高的核酸扩增检测（NAAT）对于确诊螺旋体血症阶段的早期莱姆病具有重要价值。
对有莱姆病临床表现的患者采集静脉血，进行莱姆病的标准两层筛查和免疫印迹血清学检测，同时采用巢式聚合酶链反应（PCR）检测伯氏疏螺旋体复合群16S核糖体DNA。对PCR扩增子进行测序以验证伯氏疏螺旋体基因组DNA。共研究了130名前往急诊室（ER）或随诊诊所（WALKIN）就诊的患者以及333名通过私人医生办公室转诊的患者。急诊室/随诊诊所患者中有5.4%显示有螺旋体血症的DNA证据，而私人医生办公室转诊的患者中无一例（0%）DNA呈阳性。相比之下，私人医生办公室转诊的患者中有8.4%两层莱姆血清学检测呈阳性，而急诊室/随诊诊所患者中只有1.5%该抗体检测呈阳性。两层血清学检测漏诊了85.7%的早期螺旋体血症莱姆病病例。后者的诊断通过DNA测序得以证实。
巢式PCR随后进行自动化DNA测序是标准两层抗体检测在诊断早期螺旋体血症莱姆病中的重要补充。检测莱姆螺旋体血症的最佳时机是居住在莱姆病流行区的患者在病程早期出现无法解释的症状或与莱姆病相符的临床表现时。
