ELISA with Recombinant rKRP42 Antigen Using Urine Samples: A Tool for Predicting Clinical Visceral Leishmaniasis Cases and Its Outbreak

Mohammad Zahidul Islam Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Makoto Itoh Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Md. Anwar Ul Islam Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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A. R. M. Saifuddin Ekram Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Md. Ajijur Rahman Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Hidekazu Takagi Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Atsuhide Takesue Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Yoshihisa Hashiguchi Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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Eisaku Kimura Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan; Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh; Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh; Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador; Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Japan

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We reported a highly sensitive and specific enzyme-linked immunosorbent assay (ELISA) that detects immunoglobulin G (IgG) in urine using rKRP42 antigen for the diagnosis of visceral leishmaniasis (VL). The ELISA was applied to study chronological change in antibody titers in five study areas in Rajshahi district, Bangladesh. A total of 585 subjects without a past VL history were examined at least three times in the 30-month follow-up period; of these subjects, 137 (23.4%) subjects became ELISA-positive at least one time during the study. Among the positive cases, 40 (29.2%) subjects developed clinical VL, and 31 (77.5%) of these subjects showed IgG titers of ≥ 1,000 U more than one time in the study period. Considering only the first ELISA results, 22 subjects with IgG titers of ≥ 1,000 U could be found, and 21 (95.5%) of these subjects turned out to be clinical cases. The high urinary IgG titers (≥ 1,000 U) will help predict possible clinical VL cases and thus, identify an outbreak in its earlier stage.

Author Notes

* Address correspondence to Makoto Itoh, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-ken 480-1195, Japan. E-mail: macitoh@aichi-med-u.ac.jp

Financial support: This study was supported by Japan Society for the Promotion of Science (JSPS) Grant-in-Aid for Scientific Research for JSPS Postdoctoral Fellowship for Foreign Researchers 1604227, Grant-in-Aid for Scientific Research (B) 15406018, and Grant-in-Aid for Scientific Research (B) 18406013. This work was also supported by the Special Coordination Funds for Promoting Science and Technology of Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan (1200015).

Authors' addresses: Mohammad Zahidul Islam, Makoto Itoh, Hidekazu Takagi, Atsuhide Takesue, and Eisaku Kimura, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-ken, Japan, E-mails: islammz@yahoo.com, macitoh@aichi-med-u.ac.jp, htakagi@aichi-med-u.ac.jp, atakesue@aichi-med-u.ac.jp, and kimura@aichi-med-u.ac.jp. Present address of Mohammad Zahidul Islam, Department of Biology, The Catholic University of America, Washington, DC. Md. Anwar Ul Islam and Md. Ajijur Rahman, Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh, E-mails: profanwarulislam@yahoo.com and shamim_ru2003@yahoo.com. A. R. M. Saifuddin Ekram, Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh, E-mail: armsekram@yahoo.com. Yoshihisa Hashiguchi, Prometeo Project, Centro de Biomedicina, Universidad Central del Ecuador, Quito, Ecuador and Department of Parasitology, Kochi Medical School, Kochi University, Nankoku City, Kochi, Japan, E-mail: hasiguti@kochi-u.ac.jp.

Reprint requests: Makoto Itoh, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-ken 480-1195, Japan, E-mail: macitoh@aichi-med-u.ac.jp.

  • 1.

    Trouiller P, Olliaro P, Torreele E, Orbinski J, Laing R, Ford N, 2002. Drug development for neglected diseases: a deficient market and a public health policy failure. Lancet 359: 21882194.

    • Search Google Scholar
    • Export Citation
  • 2.

    Pearson R, Sausa QA, 1996. Clinical spectrum of leishmaniasis. Clin Infect Dis 22: 113.

  • 3.

    Desjeux P, 2004. Leishmaniasis: current situation and new perspectives. Comp Immunol Microbiol Infect Dis 27: 305318.

  • 4.

    Bern C, Chowdhury R, 2006. The epidemiology of visceral leishmaniasis in Bangladesh: prospects for improved control. Indian J Med Res 123: 275288.

    • Search Google Scholar
    • Export Citation
  • 5.

    Chowdhur MA, Rafiqueuddin AK, Hossain A, 1992. Aldehyde test (formol-gel test) in the diagnosis of kala-azar (visceral leishmaniasis). Trop Doct 22: 185186.

    • Search Google Scholar
    • Export Citation
  • 6.

    Sundar S, Rai M, 2002. Laboratory diagnosis of visceral leishmaniasis. Clin Diagn Lab Immunol 9: 951958.

  • 7.

    Zijlstra EE, Ali MS, el-Hassan AM, el-Toum IA, Satti M, Ghalib HW, Kager PA, 1992. Kala-azar: a comparative study of parasitological methods and the direct agglutination test in diagnosis. Trans R Soc Trop Med Hyg 86: 505507.

    • Search Google Scholar
    • Export Citation
  • 8.

    Ghose AC, Haldar JP, Pal SC, Mishra BP, Mishra KK, 1980. Serological investigations on Indian kala-azar. Clin Exp Immunol 40: 318326.

  • 9.

    Kaul P, Malla N, Kaur S, Mahajan RC, Ganguly NK, 2000. Evaluation of a 200-kDa amastigote-specific antigen of L. donovani by enzyme-linked immunosorbent assay (ELISA) for the diagnosis of visceral leishmaniasis. Trans R Soc Trop Med Hyg 94: 173175.

    • Search Google Scholar
    • Export Citation
  • 10.

    Raj VS, Ghosh A, Dole VS, Madhubala R, Myler PJ, Stuart KD, 1999. Serodiagnosis of leishmaniasis with recombinant ORFF antigen. Am J Trop Med Hyg 61: 482487.

    • Search Google Scholar
    • Export Citation
  • 11.

    Zijlstra EE, Daifalla NS, Kager PA, Khalil EAG, Hassan AME, Reed SG, Ghalib HW, 1998. rK39 enzyme-linked immunosorbent assay for diagnosis of Leishmania donovani infection. Clin Diagn Lab Immunol 5: 717720.

    • Search Google Scholar
    • Export Citation
  • 12.

    Harith AE, Kolk AHJ, Leeuwenburg J, Muigai R, Kiugu S, Kiugu S, Laarman JJ, 1986. A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis. Trans R Soc Trop Med Hyg 80: 583587.

    • Search Google Scholar
    • Export Citation
  • 13.

    Harith AE, Kolk AHJ, Leeuwenburg J, Muigai R, Huigen E, Jelsma T, Kager PA, 1988. Improvement of a direct agglutination test for field studies of visceral leishmaniasis. J Clin Microbiol 26: 13211325.

    • Search Google Scholar
    • Export Citation
  • 14.

    Burns JM, Shreffler WG, Benson DR, Ghalib HW, Badaro R, Reed SG, 1993. Molecular characterization of a kinesin-related antigen of Leishmania chagasi that detects specific antibody in African American visceral leishmaniasis. Proc Natl Acad Sci USA 90: 775779.

    • Search Google Scholar
    • Export Citation
  • 15.

    Maalej IA, Chenik M, Louzir H, Salah AB, Bahloul C, Amri F, Dellagi K, 2003. Comparative evaluation of ELISAs based on ten recombinant or purified leishmania antigens for the serodiagnosis of Mediterranean visceral leishmaniasis. Am J Trop Med Hyg 68: 312320.

    • Search Google Scholar
    • Export Citation
  • 16.

    Islam MZ, Itoh M, Takagi H, Islam AU, Ekram ARMS, Rahman A, Takesue A, Hashiguchi Y, Kimura E, 2008. Enzyme-linked immunosorbent assay to detect urinary antibody against recombinant rKRP42 antigen made from Leishmania donovani for the diagnosis of visceral leishmaniasis. Am J Trop Med Hyg 79: 599604.

    • Search Google Scholar
    • Export Citation
  • 17.

    CDC, 2010. CDC—Parasites—Leishmaniasis. Resources for Health Professionals. Available at: http://www.cdc.gov/parasites/leishmaniasis/health_professionals/index.html. Accessed December 28, 2011.

    • Search Google Scholar
    • Export Citation
  • 18.

    Ahluwalia IB, Bern C, Costa C, Akter T, Chowdhury R, Ali M, Alam D, Kenah E, Amann J, Islam M, Wagatsuma Y, Haque R, Breiman RF, Maguire JH, 2003. Visceral leishmaniasis: consequences of a neglected disease in a Bangladeshi community. Am J Trop Med Hyg 69: 624628.

    • Search Google Scholar
    • Export Citation
  • 19.

    Collin S, Davidson R, Ritmeijer K, Keus K, Melaku Y, Kipngetich S, Davies C, 2004. Conflict and kala-azar: determinants of adverse outcome of kala-azar among patients in Southern Sudan. Clin Infect Dis 38: 612619.

    • Search Google Scholar
    • Export Citation
  • 20.

    Singh S, Kumari V, Singh N, 2002. Predicting kala-azar disease manifestations in asymptomatic patients with latent Leishmania donovani infection by detection of antibody against recombinant rK39 antigen. Clin Diagn Lab Immunol 9: 568572.

    • Search Google Scholar
    • Export Citation
  • 21.

    Islam MZ, Itoh M, Shamsuzzaman SM, Mirza R, Matin F, Ahmed I, Choudhury AKMS, Hossain MA, Qiu XG, Begam N, Furuya M, Leafasia JL, Hashiguchi Y, Kimura E, 2002. Diagnosis of visceral leishmaniasis by ELISA using urine samples. Clin Diagn Lab Immunol 9: 789794.

    • Search Google Scholar
    • Export Citation
  • 22.

    Islam MZ, Itoh M, Mirza R, Ahmed I, Ekram ARMS, Sarder AH, Shamsuzzaman SM, Hashiguchi Y, Kimura E, 2004. Direct agglutination test with urine samples for the diagnosis of visceral leishmaniasis. Am J Trop Med Hyg 70: 7882.

    • Search Google Scholar
    • Export Citation
  • 23.

    Attar ZJ, Chance ML, el-Safi S, Carney J, Azazy A, El-Hadi M, Dourado C, Hommel M, 2001. Latex agglutination test for the detection of urinary antigens in visceral leishmaniasis. Acta Trop 78: 1116.

    • Search Google Scholar
    • Export Citation
  • 24.

    Itoh M, Weerasooriya MV, Qui X-G, Gunawardena NK, Anantaphruti MT, Tesana S, Rattanaxay P, Fujimaki Y, Kimura E, 2001. Sensitive and specific enzyme-linked immunosorbent assay for the diagnosis of Wuchereria bancrofti infection in urine samples. Am J Trop Med Hyg 65: 362365.

    • Search Google Scholar
    • Export Citation
  • 25.

    Takagi H, Islam MZ, Itoh M, Islam MAU, Ekram ARMS, Hussain HM, Hashiguchi Y, Kimura E, 2007. Production of recombinant kinesin-related protein of Leishmania donovani and its application in the serodiagnosis of visceral leishmaniasis. Am J Trop Med Hyg 76: 902905.

    • Search Google Scholar
    • Export Citation
  • 26.

    Ahmed BN, 2004. Report on Outbreak Investigation, Kala azar in Godagari. IEDCR in Action, Series 2. Dhaka, Bangladesh: Institute of Epidemiology, Disease Control & Research.

    • Search Google Scholar
    • Export Citation
  • 27.

    Singh S, Reed SG, Sacks AG, Chang KP, 1995. Diagnostic and prognostic value of rK39 antigen in Indian leishmaniasis. J Parasitol 81: 10001003.

    • Search Google Scholar
    • Export Citation
  • 28.

    Weerasooriya MV, Itoh M, Islam MZ, Qiu XG, Fujimaki Y, Kimura E, 2003. Prevalence and levels of filaria-specific urinary IgG4 among children less than five years of age and the association of the antibody positivity between the children and their mothers. Am J Trop Med Hyg 68: 465468.

    • Search Google Scholar
    • Export Citation
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