Serodiagnosis of strongyloidiasis in a low-endemic setting – a two-tiered test approach

Abstract

Background

Previous studies have demonstrated that Strongyloides ELISAs frequently exhibit cross-reactivity with infections caused by other helminths. This may result in false-positive results, especially in low-endemic areas, potentially leading to unwarranted treatment. This study aimed to present a new Western blot for the detection of anti-Strongyloides antibodies such as to enhance the accuracy of distinguishing true-positive from false-positive ELISA results.

Methods

We developed and validated a Western blot to detect anti-S. stercoralis specific antibodies using reference sera from confirmed cases (n = 55), non-infected control patients (n = 20), and patients with other helminth infections (n = 50). Using this sample set, we evaluated the analytical performance of three serologic approaches: an in-house ELISA, the commercially available Bordier-ELISA, and a two-tiered testing procedure combining the in-house ELISA with a confirmatory Western blot. Using receiver operating characteristics (ROC) analysis, setting-specific cut-offs values were determined for both ELISA assays. Further, we conducted a retrospective cohort study using serum samples from solid organ transplant (SOT) candidates (n = 310) tested by ELISA during pre-transplant evaluations at Bern University Hospital (2018–2022).

Results

The Western blot specifically detected human IgG antibodies against S. stercoralis. A two-tier test algorithm (in-house ELISA, followed by confirmatory Western blot) showed a diagnostic sensitivity of 96.4 % (95 %CI:87.5 %–99.6 %) and a diagnostic specificity of 98.6 % (95 %CI:92.3 %–100 %). Out of 310 SOT candidates’ serum samples, 9.3 % (n = 29) exhibited positive, and 5.2 % (n = 16) equivocal ELISA results. Testing by Western blot revealed positive S. stercoralis findings for 11.1 % (n = 5) of these patients. Two of those were initially positive and 3 were equivocal by ELISA.

Conclusions

The new Western blot is a promising tool for clarifying equivocal S. stercoralis ELISA results, and therefore predisposed to reduce false positive Strongyloides ELISA results, especially in low-endemic regions.