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Pathogen-specific leptospiral proteins in urine of patients with febrile illness aids in differential diagnosis of leptospirosis from dengue

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Abstract

Leptospirosis and dengue are two commonly seen infectious diseases of the tropics. Differential diagnosis of leptospirosis from dengue fever is often difficult due to overlapping clinical symptoms and lack of economically viable and easy-to-perform laboratory tests. The gold standard for diagnosis is the microscopic agglutination test (MAT). In this study, the diagnostic potential of screening for pathogen-specific leptospiral antigens in urine samples is presented as a non-invasive method of disease diagnosis. In a study group of 40 patients, the serum was tested for anti-leptospiral antibodies by MAT and enzyme-linked immunosorbent assay (ELISA). Urine of these patients was screened for leptospiral antigens by ELISA using specific antibodies against LipL32, LipL41, Fla1, HbpA and sphingomyelinase. Group I patients (n = 23) were classified as leptospirosis-positive based on MAT and high titres of circulating IgM-specific anti-leptospiral antibodies. All of these patients excreted all five leptospiral antigens in the urine. The 17 MAT-negative cases included six patients with pyrexia of unknown origin (PUO; Group II) and 11 confirmed dengue patients (Group III). The latter tested negative for both serum anti-leptospiral antibodies and urinary leptospiral antigens. A salient outcome of this study was highlighting the usefulness of screening for urinary leptospiral antigens in disease diagnosis, as their presence confirmed leptospiral aetiology in two PUO patients. Immunoblots of urinary antigens identified well-defined bands corresponding to LipL32, HbpA and sphingomyelinase; the significance of the 42- and 58-kDa sphingomyelinase bands is discussed.

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Acknowledgements

KCT, CKE, BJZ and RP would like to thank the management of MOSC Medical College and the Department of Biochemistry for facilitating this study. RC and MS acknowledge the University of Hyderabad for providing the basic infrastructural facilities.

Funding

The study was not funded by any funding agency. Routine maintenance funds given to MOSC Medical College Hospital (for clinical components) and University of Hyderabad (for experimental studies) were used for the study.

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Authors and Affiliations

  1. Department of Animal Biology, School of Life Sciences, University of Hyderabad, Prof. CR Rao Road, Gachibowli, Hyderabad, 500 046, India

    R. Chaurasia & M. Sritharan

  2. Departments of Biochemistry and Medicine, MOSC Medical College, Kerala, India

    K. C. Thresiamma, C. K. Eapen, B. J. Zachariah & R. Paul

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  1. R. Chaurasia
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  2. K. C. Thresiamma
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  3. C. K. Eapen
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  4. B. J. Zachariah
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  5. R. Paul
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  6. M. Sritharan
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Contributions

MS conceptualised the study, designed the experiments and prepared the manuscript; RC performed the experiments and compiled the data; KCT coordinated the processing of the clinical samples and biochemical analyses; CKE, BJZ and RP were the clinical partners and coordinated patient selection, sample collection and collection of clinical data.

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Correspondence to M. Sritharan.

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All authors report no conflict of interest with reference to this article.

Ethical approval

Ethical clearance and informed consent from the study subjects were obtained from MOSC Medical College Hospital. Clearance from both the Institutional Ethical Committee and Institute Biosafety Committee from the University of Hyderabad were obtained for the study.

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Informed consent from the study subjects were obtained as per the guidelines of the Ethical Clearance Committee of MOSC Medical College Hospital.

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Chaurasia, R., Thresiamma, K.C., Eapen, C.K. et al. Pathogen-specific leptospiral proteins in urine of patients with febrile illness aids in differential diagnosis of leptospirosis from dengue. Eur J Clin Microbiol Infect Dis 37, 423–433 (2018). https://doi.org/10.1007/s10096-018-3187-9

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