DOI: http://dx.doi.org/10.18203/2320-6012.ijrms20170168

Proteinuria in early detection of human leptospirosis

Thresiamma K. C., Asha Biju, Reetika Chaurasia, Manjula Sritharan, Chithra Jayaprakash, Sunil Thomas, Eapen C. K.

Abstract


Background: Leptospirosis is an infectious disease caused by spirochetes bacteria Leptospira spp. and is reported from all over the world. As the clinical signs and symptoms of Leptospirosis often are nonspecific and the disease is early mistaken for other major infectious febrile illness, laboratory test to confirm the clinical diagnosis thus is essential for optimal treatment and patient management.

Methods: Serum and urine samples were collected from patients clinically suspected cases of Leptospirosis. Preparations of urine concentrate by precipitation and centrifugation.

Results: It was interesting to note that immunoglobulins are present in the urine protein concentrate of patients with Leptospirosis on the day of admission in the hospital, with urine albumin reports either positive or negative. By ELISA test it was noted that antibodies present in urine and serum were of both IgM and IgG class against the Leptospiral antigens from three pathogenic serovars and one non-pathogenicserovars. In the immunospot test which was done and compared with standard ELISA test for serum antibodies using same antigen showed that antibodies present in urine protein concentrate, which was collected on the day of admission when patients come with suspecting symptoms of Leptospirosis.

Conclusions: Proteinuria is the most frequent abnormality noted in all patients at some stage of illness. This is the first report on the presence of immunoglobulins in urine samples, which were found to be of IgM and IgG classes. These findings are of significant diagnostic potential as a simple immune-spot test can be done for detecting anti-leptospiral antibodies in urine samples of suspected cases. The present attempt was aimed at developing an immunospot test, a simple and rapid diagnostic test to detect Leptospirosis using urine samples of clinically suspected patients of the infection at the earliest. It was found to be in good correlation with standard ELISA method which is being used to detect serum antibodies in Leptospira infected patients using the same antigen.

Keywords


Immunospot, Leptospirosis, Urine IgM and IgG

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References


Douglin CP, Jordan C, Rock R, Hurley A, Levett PN. Risk factors for severe leptospirosis in parish of St. Andrew , Barbados. Emerg Infect Dis.1997;3:78-80.

Faine S. Guidelines for the control of leptospirosis. World Health Organization, Geneva, Switzerland. 1982.

Mohanan AM, Callanan JJ, Nally JE. Review paper: Host- Pathogen interactions in the Kidney during Chronic Leptospirosis. Vet Pathol. 2009;46:792-9.

Senthilkumar T, Subathre M, Phil M, Ramadass P, Ramaswamy V. Rapid diagnosis of Leptospirosis by latex agglutination test and flow-through assay. Indian. J Med Microbiology. 2008;26:45-9.

Gussenhoven GC, Van der Hoorm MA, Goris MG, Terpstra WJ, Hartskeerl RA, Mol BW, et al. LEPTO dipstick, a dipstick assay for detection of Leptospira-specific immunoglobulin M. antibodies in human sera. J Clin Microbiol. 1997;35:92-7.

Wagenaar J, Zuerner RL, Alt D, Bolin CA, Coparison of polymerase chain reaction assays with bacteriologic culture, immune-fluorescence, and nucleic acid hybridization for detection of Leptospira borgpetersenii serovar Hardjo in urine of cattle. Am. J Vet Res. 2000:61:316-20.

Croda J, Joao GR, Matsunaga J, Queiroz A, Homma A, Riley LW, et al. Leptospira Immunoglobulin-Like Proteins as a Serodiagnostic Marker for Acute Leptospirosis. J Cli Microbol. 2007:45:1528-34.

Arimitsu Y, Kmety E, Anayina Y, Baranton G, Ferguson IR, Smythe L, et al. Evaluation of the one point microcapsule agglutination test for the diagnosis of leptospirosis. Bull WHO. 1994:72:393-9.

Alder B, Murphy AM, Locarnini SA, Faine S. Detection of Specific anti-leptospiral immunoglobulins M and G in human serum solid phase enzyme- linked immunosorbent assay. J.Clin. Microbiol. 1980;11:452-7.

Milner AR, Jackson KB, Woodruff K, Smart IL. Enzyme-linked immunosorbent assay for determining specific immunoglobulin M in infections caused by Leptospira interrogans serovar hardjo. J Clin. Microbiol. 1985;22:539-42.

Pappas MG, Ballou WR, Gray MR, Takafuji ET, Miller RN, Hockmeyer WT. Rapid serodiagnosis of leptospirosis using the IgM-specific dot-ELISA: Comparison with the microscopic agglutination test. Am J Trop Med Hyg. 1985;34:346-54.

Saengjaruk P, Chaicumpa W, Watt G, Bunyaraksyotin G, Wuthiekanun V, Tapchaisri P, et al. Diagnosis of Human Leptospirosis by Monoclonal Antibody –based Antigen Detection in Urine. Clin Microbiol. 2002;40:480-9.

Widiyanti D, Koizumi N, Fukui T, Muslich LT, Segawa T, Villanueva SYAM, et al. Development of Immunochromatography-Based methods for Detection of Leptospiral Lipopolysaccharide Antigen in Urine. Clin. Vaccine Immunol. 2013;20:683-90.

Tiselius A. ‘New Approach for Electrophoretic Analysis of Colloidal Mixtures’ Trans. Faraday Soc. 1937;33:524.

Burtiz C, Edward RA, Tietz. Fundamentals of clinical chemistry. 2001;5:181-3.

Faine S. Leptospira and Leptospirosis. CRC Press Inc Boca Raton, Florida. 1994.

Faine S. AlderB, Bolin C. Perolat P: Leptospira and Leptospirosis,2nd ed, MediSci. Melbourne. Australia, 1999.

Yang CW, Wu MS, Pan MJ. Leptospirosis renal disease. Nephrol Dial Transplant. 2001;16(suppl 5):73-7.

Eapen CK, Tresiamma KC, Thomas S. Protienuria in early Human Leptospirosis used for Simple Cheap test for Leptospirosis. 2015 Abstracts , 9th Scientific meeting of International Leptospirosis Society : 46.