Lyme neuroborreliosis in Swedish children—PCR as a complementary diagnostic method for detection of Borrelia burgdorferi sensu lato in cerebrospinal fluid

Abstract

The aim of this study was to evaluate polymerase chain reaction (PCR) as a diagnostic method for the detection of Borrelia burgdorferi s.l. in CSF of Swedish children with LNB. This study was performed retrospectively on CSF and serum samples collected from children evaluated for LNB (n = 233) and controls with other specific neurological disorders (n = 59) in a Swedish Lyme endemic area. For anti-Borrelia antibody index, the IDEIA Lyme Neuroborreliosis kit (Oxoid) was used. Two in-house real-time PCR assays targeting the 16S rRNA gene were evaluated (TaqMan® and LUX™). Among patients classified as LNB cases (n = 102), five children (5%) were Borrelia PCR-positive in CSF with the TaqMan® assay. In the Non-LNB group (n = 131), one patient was Borrelia PCR positive with the TaqMan® assay. Among controls (n = 59), all CSF samples were PCR negative. When amplifying and sequencing ospA, we found B. garinii (n = 2), B. afzelii (n = 2), B. bavariensis (n = 1), and one untypable (n = 1). With the LUX™ technology, all CSF samples were PCR negative. The TaqMan® assay could detect only few cases (n = 6) of B. burgdorferi s.l. in CSF among children with LNB and the sensitivity was very low (5%). However, using larger CSF volumes and centrifugation of samples, the PCR technique could still be useful as a complementary diagnostic method when evaluating LNB. Furthermore, detection of spirochete DNA in clinical matrices, including CSF, is the method of choice for studying epidemiological aspects of LNB, a tick-borne emerging disease.

Establishment of a digital PCR method for detection of Borrelia burgdorferi sensu lato complex DNA in cerebrospinal fluid

“…We included fifty-nine patients examined for LNB, with samples stored at − 20 °C containing at least 0.5 mL CSF sample material. We detected Borrelia DNA in one clinical sample from a patient diagnosed with LNB resulting in a clinical sensitivity of 11% (1/9), which is within range of previous reports 9,10,[23][24][25] . Positive PCR results for detection of Borrelia targets in CSF samples often have high cycle threshold values (> 38), indicating low amounts of targets in the samples 10,26 .…”

“…In general, the analytical sensitivity and specificity of PCR assays detecting Borrelia in CSF samples are high 9 , 26 , 27 , but the clinical sensitivity on CSF samples is suboptimal (median sensitivity of 40%) 25 . Consequently, it has been suggested that larger CSF sample volumes (> 1 mL) in combination with centrifugation may improve the possibility of detecting the Borrelia target by PCR 8 , 10 , 20 , 23 . A different approach enabling enrichment of Borrelia DNA prior to PCR could be to culture the clinical CSF samples.…”

PathoDNA, an Advanced Diagnostic for Lyme Disease & Co-Infections Utilizing Next Generation DNA Sequencing with Greater Sensitivity and Selectivity Than ELISA/Western Blot

“…With the performance of ELISA and Western blot both lackluster in detecting Lyme disease, it is about time patients and physicians have improved technology. [68]. The tests were retrospective, allowing the researchers to categorize the children into groups of "Definite," "Possible," and "non-Lyme" in terms of their Lyme disease status; another group of children not exposed to Lyme disease was used as a control group.…”

“…The tests were retrospective, allowing the researchers to categorize the children into groups of "Definite," "Possible," and "non-Lyme" in terms of their Lyme disease status; another group of children not exposed to Lyme disease was used as a control group. The PCR tests identified only a small portion of the definite Lyme cases as positive for Lyme disease (5%) while changing one child from the "Possible" group to "Definite" and another child from the "non-Lyme" group to "Definite" [68]. The conclusion that Skogman and her colleagues drew from the results was that these two PCR tests were better suited to a complementary role in Lyme disease testing due to its very low sensitivity but could still remain useful for its ability to catch certain cases that would otherwise not be classified as a Lyme infection.…”

Epizootiological aspects of natural nidality of Ixodes tick-borne borreliosis in the Moscow region (Russian Federation)

“…Currently, Ixodes tick-borne borreliosis is widespread in the United States, Canada, Russia, and some Western European countries [3,4]. For example, cases are reported from the North West District to the Far East territories in Russia, including Sakhalin [5].…”