Lyme Borreliosis - the Utility of Improved Real-Time PCR Assay in Detection of Borrelia burgdorferi Infections

Abstract

Background. Infections of Borrelia burgdorferi sensu lato reveal clinical manifestations affecting numerous organs and tissues. The standard diagnostic procedure of these infections is quite simple if a positive history of tick exposure or typical erythema migrans appears. Lack of unequivocal clinical symptoms creates the necessity for further evaluation with laboratory tests. Objectives. This study discusses the utility of a novel, improved, well-optimized, sensitive and highly specific quantitative real-time PCR assay for the diagnostics of infections caused by Borrelia burgdorferi sensu lato. Material and Methods. We designed an improved, specific, highly sensitive real-time quantitative polymerase chain reaction (RQ-PCR) assay for the detection and quantification of all Borrelia burgdorferi genotypes. A wide validation effort was undertaken to ensure confidence in the highly sensitive and specific detection of B. burgdorferi. Results. Due to high sensitivity and great specificity, as low as 1.6 × 10² copies of Borrelia per mL of whole blood could be detected. As much as 12 (3%) negative ELISA IgM results, 14 (2.8%) negative results of Line blot IgM, 11 (3.1%) and 7 (2.7%) of negative ELISA IgG and Line blot IgG results, respectively, were positive in real-time PCR. Conclusions. The data in this study confirms the high positive predictive value of real-time PCR test in the detection of Borrelia infections (Adv Clin Exp Med 2015, 24, 4, 663-670).

Borrelia mayonii sp. nov., a member of the Borrelia burgdorferi sensu lato complex, detected in patients and ticks in the upper midwestern United States

“…Patients presented with somewhat differing clinical presentations when compared with patients infected with B. burgdorferi sensu stricto including diffuse macular rash not typical of erythema migrans (EM), nausea or vomiting and symptoms potentially consistent with neurological involvement. By oppA2 quantitative PCR and dark-field microscopy, the genome and spirochete load, in blood of acutely ill patients, was estimated to be approximately 10 5 ml −1 , which is higher than previously reported among patients infected with other Bbsl genospecies (Bil-Lula et al, 2015; Liveris et al, 2012). Motile spirochetes were cultured from blood of two patients under microaerophilic conditions at 34 °C in modified Barbour–Stoenner–Kelly (BSK) medium.…”

The role of companion animals in the environmental circulation of tick-borne bacterial pathogens

“…Some genes of Bbsl are expressed only in a mammalian host or have significantly upregulated expression in that environment, for example, VlsE, DbpA, BBK32, Erp, and Mlp proteins. For these reasons the detection of bacteria genomic DNA may play an important role in the diagnosis of Borrelia infections [70][71][72].…”

Whole Genome Sequence and Comparative Genomics of the Novel Lyme Borreliosis Causing Pathogen, Borrelia mayonii

“…mayonii infected patients were shown to have elevated numbers of spirochetes/genomes in their blood (10 5 to 10 6 spirochetes/genomes per milliliter) as compared to loads previously reported for B . burgdorferi (10 2 to 10 3 genomes per milliliter; 0.1 spirochete per milliliter) [14–16]. This elevated spirochetemia coupled with blood based PCR detection for five patients led to the suggestion that B .…”