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Enzyme-linked Immunosorbent Assay to Detect Urinary Antibody Against Recombinant rKRP42 Antigen Made from Leishmania donovani for the Diagnosis of Visceral Leishmaniasis

We recently reported the production of the recombinant kinesin-related protein of Leishmania donovani with a molecular weight of 42 kd (rKRP42) and the value of the antigen in serum-based ELISA for the diagnosis of visceral leishmaniasis (VL). In this study, the rKRP42 antigen was validated with ELISA using urine samples (rKRP42 urine ELISA). The urine-based ELISA showed 94% sensitivity (108 positives among 115 VL samples) and 99.6% specificity (239 negatives among 240 non-VL samples). The sensitivity and specificity are almost similar to our previous results by ELISA with acetone-treated L. donovani promastigote antigen and direct agglutination test, both methods being done by use of urine samples. A comparison of the rKRP42 urine ELISA with the commercially available urinary antigen detection kit (KAtex) using 108 VL samples showed much higher sensitivity of the ELISA (96.3%) than KAtex (55.6%). The use of the rKRP42 antigen with urine samples will facilitate epidemiologic studies.

Received June 3, 2008. Accepted for publication July 17, 2008.

Acknowledgments: We thank Drs. Rusella Mirza, Farzana Matin, Mohsin Ali, A. K. M. Shamsuzzaman Choudhury, Akram Hossain, Nilufar Begam, and Sultana Monira Hussain for helping in collection of VL samples and Dr. Judson. L. Leafasia for providing malaria samples.

Financial support: This study was supported by Grant-in-Aid for Scientific Research for JSPS Postdoctoral Fellowship for Foreign Researchers 16. 04227, Grant-in-Aid for Scientific Research (B) 15406018, and Grant-in-Aid for Scientific Research (B) 18406013 from the Japan Society for the Promotion of Science, and also by the Special Coordination Funds for Promoting Science and Technology of MEXT, Japan, 1200015.

^* Address correspondence to Mohammad Zahidul Islam, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-ken 480-1195, Japan, 5. E-mail: islammz{at}yahoo.com

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 480-1195, Japan, Tel: 81-52-264-4811, Fax: 81-561-63-3645, E-mails: zahid{at}aichi-med-u.ac.jp, macitoh{at}aichi-med-u.ac.jp, htakagi{at}aichi-med-u.ac.jp, atakesue{at}aichi-med-u.ac.jp, and kimura{at}aichi-med-u.ac.jp. Anwar Ul Islam and Ajijur Rahman, Department of Pharmacy, University of Rajshahi, Rajshahi, Bangladesh, Tel: 88-721-750041-9, Fax: 88-721-750064, E-mails: profanwarulislam{at}yahoo.com and shamim_ru2003{at}yahoo.com. A. R. M. Saifuddin Ekram, Department of Medicine, Rajshahi Medical College, Rajshahi, Bangladesh, Tel: 88-721-776001-9, Fax: 88-721-772174, E-mail: armsekram{at}yahoo.com. Yoshihisa Hashiguchi, Department of Parasitology, Kochi Medical school, Kochi University, Nankoku City, Kochi 783-8505, Japan, Tel: 81-88-880-2415, Fax: 81-88-880-2415, E-mail: hasiguti{at}kochi-u.ac.jp.

Reprint requests: Mohammad Zahidul Islam, Department of Parasitology, Aichi Medical University School of Medicine, Nagakute, Aichi-ken 480-1195, Japan, E-mails: zahid{at}aichi-med-u.ac.jp and islammz{at}yahoo.com.