Diagnostic method-based underestimation of leptospirosis in clinical and research settings; an experience from a large prospective study in a high endemic setting

Author summaryDiagnostics of leptospirosis have not been optimised yet and is considered as a significant limiting factor for estimating the disease burden. This prospective hospital-based study, including 1734 clinically suspected leptospirosis cases and undifferentiated febrile patients, revealed that a minimum of 40% of cases would be missed by using any of the following tests individually. (Whole blood qPCR, single or paired-sample MAT, Urine qPCR, culture, culture qPCR, surveillance case definition). Therefore, we conclude that whole blood qPCR should be the standard test for diagnosing leptospirosis for clinical purposes until day 10 of the reported disease. MAT should be limited to places where the serological diagnosis has an epidemiological interest. qPCR testing of microscopically-negative cultures should be done before discarding to increase the yield in research settings. BackgroundLeptospirosis has globally significant human mortality and morbidity, yet estimating the clinical and public health burden of leptospirosis is challenging because timely diagnosis remains limited. The goal of the present study was to evaluate leptospirosis undercounting by current standard methods in both clinical and epidemiological study settings. Methodology/Principal findingsA prospective hospital-based study was conducted in multiple hospitals in Sri Lanka from 2016 to 2019. Culture, whole blood, and urine samples were collected from clinically suspected leptospirosis cases and patients with undifferentiated fever. Analysis of biological samples from 1,734 subjects confirmed 591 (34.1%) cases as leptospirosis and 297 (17.1%) were classified as probable leptospirosis cases. Whole blood quantitative PCR (qPCR) did identify the most cases (322/540(60%)) but missed 40%. Cases missed by each method include; urine qPCR, 70% (153/220); acute sample microscopic agglutination test (MAT), 80% (409/510); paired serum sample MAT, 58% (98/170); and surveillance clinical case definition, 53% (265/496). qPCR of negative culture samples after six months of observation was of diagnostic value retrospectively with but missed 58% of positives (109/353). ConclusionLeptospirosis disease burden estimates should consider the limitations of standard diagnostic tests. qPCR of multiple sample types should be used as a leading standard test for diagnosing acute leptospirosis.

Automated summary

This study found that current standard diagnostic methods can easily miss cases of leptospirosis, and whole blood qPCR can be used as the leading standard test for diagnosing acute cases, while MAT should be limited to places with epidemiological interest. Additionally, qPCR testing of microscopically-negative culture samples should be done before discarding to increase the yield.