J Korean Med Sci.  2016 Feb;31(2):183-189. 10.3346/jkms.2016.31.2.183.

Clinical Evaluation of Rapid Diagnostic Test Kit Using the Polysaccharide as a Genus-Specific Diagnostic Antigen for Leptospirosis in Korea, Bulgaria, and Argentina

• Lee JW
• Park S
• Kim SH
• Christova
• Jacob P
• Vanasco NB
• Kang YM
• Woo YJ
• Kim MS
• Kim YJ
• Cho MK
• Kim YW

• KMID: 2360040
• DOI: http://doi.org/10.3346/jkms.2016.31.2.183

Abstract

Leptospirosis, a zoonotic disease that is caused by many serovars which are more than 200 in the world, is an emerging worldwide disease. Accurate and rapid diagnostic tests for leptospirosis are a critical step to diagnose the disease. There are some commercial kits available for diagnosis of leptospirosis, but the obscurity of a species- or genus-specific antigen of pathogenic Leptospira interrogans causes the reduced sensitivity and specificity. In this study, the polysaccharide derived from lipopolysaccharide (LPS) of nonpathogenic Leptospira biflexa serovar patoc was prepared, and the antigenicity was confirmed by immunoblot and enzyme linked immunosorbent assay (ELISA). The performance of the rapid diagnostic test (RDT) kit using the polysaccharide as a diagnostic antigen was evaluated in Korea, Bulgaria and Argentina. The sensitivity was 93.9%, 100%, and 81.0% and the specificity was 97.9%, 100%, and 95.4% in Korea (which is a rare region occurring with 2 serovars mostly), Bulgaria (epidemic region with 3 serovars chiefly) and Argentina (endemic region with 19 serovars mainly) respectively. These results indicate that this RDT is applicable for global diagnosis of leptospirosis. This rapid and effective diagnosis will be helpful for diagnosis and manage of leptospirosis to use and the polysaccharide of Leptospira may be called as genus specific antigen for diagnosis.

Keyword

Leptospirosis; Genus-specific Diagnostic Antigen; Rapid Diagnosis

MeSH Terms

Antigens, Bacterial/*immunology
Argentina
Bulgaria
Enzyme-Linked Immunosorbent Assay
Female
Humans
Leptospira/isolation & purification/metabolism
Leptospira interrogans/isolation & purification/metabolism
Leptospirosis/*diagnosis/microbiology
Male
Polysaccharides/*immunology
Reagent Kits, Diagnostic/*standards
Republic of Korea
Sensitivity and Specificity
Antigens, Bacterial
Polysaccharides
Reagent Kits, Diagnostic

Figure

• 

  Fig. 1 Examination of polysaccharide as common epitope for Leptospira. (A) Polysaccharide prepared from LPS of L. biflexa serovar patoc or L. interrogans serovar lai was analyzed by SDS-PAGE and silver-staining. (B) Purified antigens were transferred onto nitrocellulose (NC) membrane and 12-20 kDa of polysaccharide was detected by human pooled serum of leptospirosis patients as common antigen to both patoc and lai.

• 

  Fig. 2 Representative pictures for RDT indicating negative and positive results. Red colorization of the test line (T) indicates the presence of human antibody against Leptospira and the red control line (C) represents the valid test result. Demonstration is negative (kit 1), both positive in IgM and IgG (kit 2), IgM positive (kit 3), and IgG positive (kit 4) in RDT results.

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