Prospective comparison of two enzyme-linked immunosorbent spot assays for the diagnosis of Lyme neuroborreliosis

Abstract

Summary Commercial cellular tests are used to diagnose Lyme borreliosis (LB), but studies on their clinical validation are lacking. This study evaluated the utility of an in‐house and a commercial enzyme‐linked immunosorbent spot (ELISpot) assay for the diagnosis of Lyme neuroborreliosis (LNB). Prospectively, peripheral blood mononuclear cells (PBMCs) were isolated from patients and controls and analysed using an in‐house Borrelia ELISpot assay and the commercial LymeSpot assay. B. burgdorferi B31 whole cell lysate and a mixture of outer surface proteins were used to stimulate the PBMCs and the numbers of interferon‐gamma‐secreting T cells were measured. Results were evaluated using receiver operating characteristic (ROC) curve analysis. Eighteen active and 12 treated LNB patients, 10 healthy individuals treated for an early (mostly cutaneous) manifestation of LB in the past and 47 untreated healthy individuals were included. Both assays showed a poor diagnostic performance with sensitivities, specificities, positive and negative predictive values ranging from 44.4–66.7%, 42.0–72.5%, 21.8–33.3% and 80.5–87.0%, respectively. The LymeSpot assay performed equally poorly when the calculation method of the manufacturer was used. Both the in‐house and the LymeSpot assay are unable to diagnose active LNB or to monitor antibiotic treatment success.

The Usefulness of Two CXCL13 Assays on Cerebrospinal Fluid for the Diagnosis of Lyme Neuroborreliosis: a Retrospective Study in a Routine Clinical Setting

“…These 150 consecutive patients did not significantly differ regarding age and gender from the 1,098 consecutive patients for whom at least one LP was performed in the predefined study period (data not shown). Six additional LNB patients, who were part of two other studies from our research group ( 41 , 42 ), were selected from outside this period, to increase the number of LNB patients. In total, 78 men and 78 women were included in the study.…”

“…Due to the relatively low number of LNB patients with sufficient material in the predefined study period, additional LNB patients (definite or possible LNB) were chosen from outside the study period (from February 2011 to July 2013 and from July 2016 to November 2017). The LNB patients who were eligible had taken part in two other studies of our research group ( 41 , 42 ), and consequently, both CSF and serum from the time of diagnosis had been stored. For these additional LNB patients, the same inclusion criteria applied as for all consecutive patients.…”

“…The detection of Borrelia -specific total immunoglobulin (IgM and IgG) in serum was done by using the C6 enzyme-linked immunosorbent assay (ELISA) (Immunetics, Boston, MA, USA), and equivocal and positive C6 ELISA results were confirmed by using the recom Line IgM and IgG immunoblot test (Mikrogen Diagnostik GmbH) as described previously ( 42 ). The detection of intrathecally produced Borrelia -specific IgM and IgG was done by using the second-generation IDEIA LNB assay (Oxoid, Basingstoke, United Kingdom) as described previously ( 42 ). To determine the intrathecal synthesis of Borrelia -specific IgM (or IgG), CSF and serum of a CSF-serum pair were tested simultaneously in the same run.…”

ELISPOT assays and their diagnostic potential in Lyme disease and Lyme neuroborreliosis

“…We have read the research article entitled ‘Prospective comparison of two enzyme‐linked immunosorbent spot assays for the diagnosis of Lyme neuroborreliosis ’ by van Gorkom T et al , published in Clinical and Experimental Immunology [1]. We want to congratulate the authors for the publishing of this article, and make some remarks and contributions.…”

“…The study population of active LNB included in the study by Gorkom et al . [1] included 6 out of 18 patients with suspected LNB because of the lack of intrathecal antibodies against Borrelia. The interesting study population in this case would be the correlation of the remaining twelve patients with definite active LNB and the EliSpot result, which was missing in the results and discussion part, at least their Table 5b, which is not included but shortly mentioned in the manuscript.…”