• 1.

    Pappas G, Papadimitriou P, Siozopoulou V, Christou L, Akritidis N, 2008. The globalization of leptospirosis: worldwide incidence trends. Int J Infect Dis 12: 351357.

    • Search Google Scholar
    • Export Citation
  • 2.

    Levett PN, 2001. Leptospirosis. Clin Microbiol Rev 14: 296326.

  • 3.

    Costa F, Hagan JE, Calcagno J, Kane M, Torgerson P, Martinez-Silveira MS, Stein C, Abela-Ridder B, Ko AI, 2015. Global morbidity and mortality of leptospirosis: a systematic review. PLoS Negl Trop Dis 9: e0003898.

    • Search Google Scholar
    • Export Citation
  • 4.

    Cruz LS, Vargas R, Lopes AA, 2009. Leptospirosis: a worldwide resurgent zoonosis and important cause of acute renal failure and death in developing nations. Ethn Dis 19: 3741.

    • Search Google Scholar
    • Export Citation
  • 5.

    Sethi S, Sharma N, Kakkar N, Taneja J, Chatterjee SS, Banga SS, Sharma M, 2010. Increasing trends of leptospirosis in northern India: a clinico-epidemiological study. PLoS Negl Trop Dis 4: 579.

    • Search Google Scholar
    • Export Citation
  • 6.

    Victoriano AF et al. 2009. Leptospirosis in the Asia Pacific region. BMC Infect Dis 9: 147.

  • 7.

    Lim VK, 2011. Leptospirosis: a re-emerging infection. Malays J Pathol 33: 15.

  • 8.

    Reller ME, Bodinayake C, Nagahawatte A, Devasiri V, Kodikara-Arachichi W, Strouse JJ, Flom JE, Dumler JS, Woods CW, 2011. Leptospirosis as frequent cause of acute febrile illness in southern Sri Lanka. Emerg Infect Dis 17: 16781684.

    • Search Google Scholar
    • Export Citation
  • 9.

    Bourhy P et al. 2012. Human Leptospira isolates circulating in Mayotte (Indian Ocean) have unique serological and molecular features. J Clin Microbiol 50: 307311.

    • Search Google Scholar
    • Export Citation
  • 10.

    Fonzar UJ, Langoni H, 2012. Geographic analysis on the occurrence of human and canine leptospirosis in the city of Maringá, state of Paraná, Brazil. Rev Soc Bras Med Trop 45: 100105.

    • Search Google Scholar
    • Export Citation
  • 11.

    Lau CL, Dobson AJ, Smythe LD, Fearnley EJ, Skelly C, Clements AC, Craig SB, Fuimaono SD, Weinstein P, 2012. Leptospirosis in American Samoa 2010: epidemiology, environmental drivers, and the management of emergence. Am J Trop Med Hyg 86: 309319.

    • Search Google Scholar
    • Export Citation
  • 12.

    Zhang C, Wang H, Yan J, 2012. Leptospirosis prevalence in Chinese populations in the last two decades. Microbes Infect 14: 317323.

  • 13.

    McBride AJ, Athanazio DA, Reis MG, Ko AI, 2005. Leptospirosis. Curr Opin Infect Dis 18: 376386.

  • 14.

    Ko AI, Goarant C, Picardeau M, 2009. Leptospira: the dawn of the molecular genetics era for an emerging zoonotic pathogen. Nat Rev Microbiol 7: 736747.

    • Search Google Scholar
    • Export Citation
  • 15.

    Zaki SR, Shieh WJ, 1996. Leptospirosis associated with outbreak of acute febrile illness and pulmonary haemorrhage, Nicaragua, 1995. The epidemic working group at ministry of health in Nicaragua. Lancet 347: 535536.

    • Search Google Scholar
    • Export Citation
  • 16.

    Gouveia EL, Metcalfe J, de Carvalho AL, Aires TS, Villasboas-Bisneto JC, Queirroz A, Santos AC, Salgado K, Reis MG, Ko AI, 2008. Leptospirosis-associated severe pulmonary hemorrhagic syndrome, Salvador, Brazil. Emerg Infect Dis 14: 505508.

    • Search Google Scholar
    • Export Citation
  • 17.

    Papa A, Theoharidou D, Antoniadis A, 2009. Pulmonary involvement and leptospirosis, Greece. Emerg Infect Dis 15: 834835.

  • 18.

    Crump JA et al. 2013. Etiology of severe non-malaria febrile illness in northern Tanzania: a prospective cohort study. PLoS Negl Trop Dis 7: e2324.

    • Search Google Scholar
    • Export Citation
  • 19.

    LaRocque RC, Breiman RF, Ari MD, Morey RE, Janan FA, Hayes JM, Hossain MA, Brooks WA, Levett PN, 2005. Leptospirosis during dengue outbreak, Bangladesh. Emerg Infect Dis 11: 766769.

    • Search Google Scholar
    • Export Citation
  • 20.

    Abela-Ridder B, Sikkema R, Hartskeerl RA, 2010. Estimating the burden of human leptospirosis. Int J Antimicrob Agents 36 (Suppl 1): S5S7.

  • 21.

    Biggs HM, Hertz JT, Munishi OM, Galloway RL, Marks F, Saganda W, Maro VP, Crump JA, 2013. Estimating leptospirosis incidence using hospital-based surveillance and a population-based health care utilization survey in Tanzania. PLoS Negl Trop Dis 7: e2589.

    • Search Google Scholar
    • Export Citation
  • 22.

    Levett PN, 2007. Leptoapira. Manual of Clinical Microbiiology, 9th edition. Washington, DC: American Society for Microbiolgy Press, 963970.

  • 23.

    Bharti AR et al. 2003. Leptospirosis: a zoonotic disease of global importance. Lancet Infect Dis 3: 757771.

  • 24.

    Flannery B et al. 2001. Evaluation of recombinant Leptospira antigen-based enzyme-linked immunosorbent assays for the serodiagnosis of leptospirosis. J Clin Microbiol 39: 33033310.

    • Search Google Scholar
    • Export Citation
  • 25.

    Srimanote P, Wongdeethai N, Jieanampunkul P, Samonkiert S, Leepiyasakulchai C, Kalambaheti T, Prachayasittikul V, 2008. Recombinant ligA for leptospirosis diagnosis and ligA among the Leptospira spp. clinical isolates. J Microbiol Methods 72: 7381.

    • Search Google Scholar
    • Export Citation
  • 26.

    Chalayon P, Chanket P, Boonchawalit T, Chattanadee S, Srimanote P, Kalambaheti T, 2011. Leptospirosis serodiagnosis by ELISA based on recombinant outer membrane protein. Trans R Soc Trop Med Hyg 105: 289297.

    • Search Google Scholar
    • Export Citation
  • 27.

    Chen HW, Zhang Z, Halsey ES, Guevara C, Canal E, Hall E, Maves R, Tilley DH, Kochel TJ, Ching WM, 2013. Detection of Leptospira-specific antibodies using a recombinant antigen-based enzyme-linked immunosorbent assay. Am J Trop Med Hyg 89: 10881094.

    • Search Google Scholar
    • Export Citation
  • 28.

    Kitashoji E et al. 2015. Diagnostic accuracy of recombinant immunoglobulin-like protein a-based IgM ELISA for the early diagnosis of leptospirosis in the Philippines. PLoS Negl Trop Dis 9: e0003879.

    • Search Google Scholar
    • Export Citation
  • 29.

    Studier FW, Moffatt BA, 1986. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol 189: 113130.

    • Search Google Scholar
    • Export Citation
  • 30.

    Matsunaga J et al. 2003. Pathogenic Leptospira species express surface-exposed proteins belonging to the bacterial immunoglobulin superfamily. Mol Microbiol 49: 929945.

    • Search Google Scholar
    • Export Citation
  • 31.

    Frey A, Di Canzio J, Zurakowski D, 1998. A statistically defined endpoint titer determination method for immunoassays. J Immunol Methods 221: 3541.

    • Search Google Scholar
    • Export Citation
  • 32.

    Joseph L, Gyorkos TW, Coupal L, 1995. Bayesian estimation of disease prevalence and the parameters of diagnostic tests in the absence of a gold standard. Am J Epidemiol 141: 263272.

    • Search Google Scholar
    • Export Citation
  • 33.

    Dendukuri N, Joseph L, 2001. Bayesian approaches to modeling the conditional dependence between multiple diagnostic tests. Biometrics 57: 158167.

    • Search Google Scholar
    • Export Citation
  • 34.

    Vacek PM, 1985. The effect of conditional dependence on the evaluation of diagnostic tests. Biometrics 41: 959968.

  • 35.

    Branscum AJ, Gardner IA, Johnson WO, 2005. Estimation of diagnostic-test sensitivity and specificity through Bayesian modeling. Prev Vet Med 68: 145163.

    • Search Google Scholar
    • Export Citation
  • 36.

    Palaniappan RU et al. 2002. Cloning and molecular characterization of an immunogenic LigA protein of Leptospira interrogans. Infect Immun 70: 59245930.

    • Search Google Scholar
    • Export Citation
  • 37.

    Koizumi N, Watanabe H, 2004. Leptospiral immunoglobulin-like proteins elicit protective immunity. Vaccine 22: 15451552.

  • 38.

    Mcbride AJ, Cerqueira GM, Suchard MA, Moreira AN, Zuerner RL, Reis MG, Haake DA, Ko AI, Dellagostin OA, 2009. Genetic diversity of the leptospiral immunoglobulin-like (Lig) genes in pathogenic Leptospira spp. Infect Genet Evol 9: 196205.

    • Search Google Scholar
    • Export Citation
  • 39.

    Agresti A, 1996. An Introduction to Categorical Data Analysis. New York, NY: Wiley.

  • 40.

    Guerreiro H, Croda J, Flannery B, Mazel M, Reis MG, Levett PN, Ko AI, Haake DA, 2001. Leptospiral proteins recognized during the humoral immune response to leptospirosis in humans. Infect Immun 69: 49584968.

    • Search Google Scholar
    • Export Citation
  • 41.

    Sugunan AP, Natarajaseenivasan K, Vijayachari P, Sehgal SC, 2004. Percutaneous exposure resulting in laboratory-acquired leptospirosis—a case report. J Med Microbiol 53: 12591262.

    • Search Google Scholar
    • Export Citation
  • 42.

    Smits HL et al. 2001. Lateral-flow assay for rapid serodiagnosis of human leptospirosis. Clin Diagn Lab Immunol 8: 166169.

  • 43.

    Effler PV, Bogard AK, Domen HY, Katz AR, Higa HY, Sasaki DM, 2002. Evaluation of eight rapid screening tests for acute leptospirosis in Hawaii. J Clin Microbiol 40: 14641469.

    • Search Google Scholar
    • Export Citation
  • 44.

    Levett PN, Branch SL, 2002. Evaluation of two enzyme-linked immunosorbent assay methods for detection of immunoglobulin M antibodies in acute leptospirosis. Am J Trop Med Hyg 66: 745748.

    • Search Google Scholar
    • Export Citation
  • 45.

    Vijayachari P, Sugunan AP, Sehgal SC, 2002. Evaluation of Lepto Dri Dot as a rapid test for the diagnosis of leptospirosis. Epidemiol Infect 129: 617621.

    • Search Google Scholar
    • Export Citation
  • 46.

    Bajani MD et al. 2003. Evaluation of four commercially available rapid serologic tests for diagnosis of leptospirosis. J Clin Microbiol 41: 803809.

    • Search Google Scholar
    • Export Citation
  • 47.

    Sehgal SC, Vijayachari P, Sugunan AP, Umapathi T, 2003. Field application of lepto lateral flow for rapid diagnosis of leptospirosis. J Med Microbiol 52: 897901.

    • Search Google Scholar
    • Export Citation
  • 48.

    McBride AJ, Athanazio DA, Reis MG, Ko AI, 2005. Leptospirosis. Curr Opin Infect Dis 18: 376386.

  • 49.

    Blacksell SD et al. 2006. Limited diagnostic capacities of two commercial assays for the detection of Leptospira immunoglobulin M antibodies in Laos. Clin Vaccine Immunol 3: 11661169.

    • Search Google Scholar
    • Export Citation
  • 50.

    Desakorn V, Wuthiekanun V, Thanachartwet V, Sahassananda D, Chierakul W, Apiwattanaporn A, Day NP, Limmathurotsakul D, Peacock SJ, 2012. Accuracy of a commercial IgM ELISA for the diagnosis of human leptospirosis in Thailand. Am J Trop Med Hyg 86: 524527.

    • Search Google Scholar
    • Export Citation
  • 51.

    Lessa-Aquino C et al. 2013. Identification of seroreactive proteins of Leptospira interrogans serovar copenhageni using a high-density protein microarray approach. PLoS Negl Trop Dis 7: e2499.

    • Search Google Scholar
    • Export Citation
  • 52.

    Cerqueira GM, McBride AJ, Picardeau M, Ribeiro SG, Moreira AN, Morel V, Reis MG, Ko AI, Dellagostin OA, 2009. Distribution of the leptospiral immunoglobulin-like (lig) genes in pathogenic Leptospira species and application of ligB to typing leptospiral isolates. J Med Microbiol 58: 11731181.

    • Search Google Scholar
    • Export Citation
  • 53.

    Haake DA, Chao G, Zuerner RL, Barnett JK, Barnett D, Mazel M, Matsunaga J, Levett PN, Bolin CA, 2000. The leptospiral major outer membrane protein LipL32 is a lipoprotein expressed during mammalian infection. Infect Immun 68: 22762285.

    • Search Google Scholar
    • Export Citation
  • 54.

    Collier WA, Mochtar A, 1939. Een serologish afwijkende leptospira-stain uit der nier eener vleermuis. Gen Tijdschr Ned Ind 79: 226.

  • 55.

    Matthias MA et al. 2008. Human leptospirosis caused by a new, antigenically unique Leptospira associated with a Rattus species reservoir in the Peruvian Amazon. PLoS Negl Trop Dis 2: e213.

    • Search Google Scholar
    • Export Citation
  • 56.

    Johnson MA et al. 2004. Environmental exposure and leptospirosis, Peru. Emerg Infect Dis 10: 10161022.

  • 57.

    Alarcón-Villaverde JO, Romani-Romani F, Tejada RA, Wong-Chero P, Céspedes-Zambrano M, 2014. Leptospirosis seroprevalence and associated features in rice farmers of tropical region of Peru. Rev Peru Med Exp Salud Publica 31: 195203.

    • Search Google Scholar
    • Export Citation
  • 58.

    Cumberland P, Everard CO, Levett PN, 1999. Assessment of the efficacy of an IgM-elisa and microscopic agglutination test (MAT) in the diagnosis of acute leptospirosis. Am J Trop Med Hyg 61: 731734.

    • Search Google Scholar
    • Export Citation
  • 59.

    Limmathurotsakul D, Turner EL, Wuthiekanun V, Thaipadungpanit J, Suputtamongkol Y, Chierakul W, Smythe LD, Day NP, Cooper B, Peacock SJ, 2012. Fool’s gold: why imperfect reference tests are undermining the evaluation of novel diagnostics: a reevaluation of 5 diagnostic tests for leptospirosis. Clin Infect Dis 55: 322331.

    • Search Google Scholar
    • Export Citation
  • 60.

    Niloofa R et al. 2015. Diagnosis of leptospirosis: comparison between microscopic agglutination test, IgM-ELISA and IgM rapid immunochromatography test. PLoS One 10: e0129236.

    • Search Google Scholar
    • Export Citation
  • 61.

    Levett PN, 2003. Usefulness of serologic analysis as a predictor of the infecting serovar in patients with severe leptospirosis. Clin Infect Dis 36: 447452.

    • Search Google Scholar
    • Export Citation
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

 

 

 

An Improved Enzyme-Linked Immunoassay for the Detection of Leptospira-Specific Antibodies

View More View Less
  • 1 Naval Medical Research Center, Silver Spring, Maryland;
  • | 2 Naval Medical Research Unit No. 6, Lima, Peru;
  • | 3 Naval Medical Center San Diego, San Diego, California;
  • | 4 Department of Statistics, University of Connecticut, Storrs, Connecticut
Restricted access

Leptospirosis is a neglected zoonotic disease with worldwide endemicity and continues to be a significant public health burden on resource-limited populations. Previously, we produced three highly purified recombinant antigens (rLipL32, rLipL41, and rLigA-Rep) and evaluated their performance of detecting Leptospira-specific antibodies in enzyme-linked immunosorbent assay (ELISA) as compared with the microscopic agglutination test (MAT). The overall sensitivity of this assay approached 90%. Recently, another recombinant antigen (rLigB-Rep) was prepared. We tested each individual antigen and a 1:1:1:1 mixture of these four antigens for the detection of Leptospira-specific antibodies in ELISA. The performance of these recombinant antigens was evaluated with a much larger febrile patient panel (337 MAT-confirmed positive sera and 92 MAT-negative sera from febrile patients). Combining the detection results of immunoglobulin M and immunoglobulin G from these four individual antigens, the overall sensitivity was close to 90% but the specificity was only 66%, based on the MAT reference method. The overall sensitivity and specificity of the four-antigen mixture were 82% and 86%, respectively. The mixture of four antigens also exhibited a broader reactivity with MAT-positive samples of 18 serovars from six major pathogenic Leptospira species. Given the limitations of MAT, the data were further analyzed by Bayesian latent class model, showing that ELISA using a 1:1:1:1 mixture still maintained high sensitivity (79%) and specificity (88%) as compared with the sensitivity (90%) and specificity (83%) of MAT. Therefore, ELISA using a mixture of these four antigens could be a very useful test for seroprevalence studies.

Author Notes

Address correspondence to Wei-Mei Ching, Naval Medical Research Center, 503 Robert Grant Ave., Silver Spring, MD 20910. E-mail: wei-mei.m.ching.civ@mail.mil

Financial support: This research was supported by Naval Medical Research Center, research work unit 6000.RAD1.J.A0310 and the United States Department of Defense Global Emerging Infections Systems Research Program, work unit number 847705.82000.25GB.B0016.

Copyright statement: The views expressed in this article are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, or the U.S. government. E. S. H, C. G., E. C., E. H., R. C. M., D. H. T., T. J. K., and W-M. C. are employees of the U.S. government. This work was prepared as part of their official duties. Title 17 U.S.C. §105 provides that “Copyright protection under this title is not available for any work of the United States Government.” Title 17 U.S.C. §101 defines a U.S. government work as a work prepared by a military service member or employee of the U.S. government as part of that person’s official duties.

Authors’ addresses: Hua-Wei Chen, Heather Lukas, Kira Becker, Giulia Weissenberger, Eric Hall, Tadeusz J. Kochel, and Wei-Mei Ching, Naval Medical Research Center, Silver Spring, MD, E-mails: huawei.chen2.ctr@mail.mil, lukas.heather@gmail.com, kbecker2015@yahoo.com, gweis990@gmail.com, eric.r.hall2.mil@mail.mil, tadeusz.kochel@nbacc.dhs.gov, and wei-mei.m.ching.civ@mail.mil. Eric S. Halsey, Carolina Guevara, and Enrique Canal, Naval Medical Research Unit No. 6, Lima, Peru, E-mails: ycw8@cdc.gov, carolina.guevara.fn@mail.mil, and enrique.a.canal.fn@mail.mil. Ryan C. Maves, Naval Medical Research Unit No. 6, Lima, Peru, and Naval Medical Center San Diego, San Diego, CA, E-mail: ryan.c.maves.mil@mail.mil. Drake H. Tilley, Naval Medical Center San Diego, San Diego, CA, E-mail: drake.h.tilley.mil@mail.mil. Lynn Kuo, Department of Statistics, University of Connecticut, Storrs, CT, E-mail: lynn.kuo@uconn.edu.

Save