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Cathepsin L1, a cysteine protease secreted by the gastrodermis of juvenile and adult Fasciola gigantica, was expressed in Escherichia coli as a calmodulin binding peptide fusion protein with a molecular mass of approximately 35 kD. The recombinant cathepsin L1 (rCTL1) was tested for its antigenic potential in a cystatin capture enzyme-linked immunosorbent assay (ELISA) to diagnose human fascioliasis. The ELISA plates were sensitized with chicken egg cystatin and incubated with bacterial lysates containing the recombinant protein before the standard ELISA procedures were performed. Analysis of the sera of 13 patients infected with F. gigantica (group 1), 204 patients with other parasitic infections (group 2), 32 cholangiocarcinoma patients (group 3), and 42 healthy controls (group 4) showed that the sensitivity, specificity, accuracy, positive predictive value, and negative predictive value of this ELISA using rCTL1 were 100%, 98.92%, 98.97%, 81.25%, and 100%, respectively. These results indicate that this assay has high sensitivity and specificity in the diagnosis of human fascioliasis. In addition, we have produced sufficient amounts of antigen for use in diagnosis.
Received April 1, 2004. Accepted for publication April 30, 2004.
Acknowledgments: We thank Dr. Nimit Morakote (Faculty of Medicine, Chiang Mai University, Chang Mai, Thailand) for providing trichinosis sera and Dr. Mark Roselieb for improving the English language presentation of the manuscript.
Financial support: This research was supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. Program (grant no. PHD/0155/2542 to Chairat Tantrawatpan) and Wanchai Malee-wong and a grant from Khon Kaen University.
Authors’ addresses: Chairat Tantrawatpan, Chaisiri Wongkham, and Sopit Wongkham, Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Wanchai Maleewong and Pewpan M. Intapan, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Kunio Nakashima, Faculty of Human Health, Tokai Gakuen University, Nakahira 2-901, Tenpaku-ku, Nagoya, Aichi, Japan.
Reprint requests: Wanchai Maleewong, Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Telephone: 66-43-348387, Fax: 66-43-202475, E-mail: wanch_ma{at}kku.ac.th.
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