title: Development of an Immunochromatographic Test for Diagnosis of Visceral Leishmaniasis Based on Detection of a Circulating Antigen.
abstract: BACKGROUND
  Visceral leishmaniasis (VL) is a life-threatening disease caused by protozoan parasites of the Leishmania donovani complex. Early case detection followed by adequate treatment is essential to the control of VL. However, the available diagnostic tests are either invasive and require considerable expertise (parasitological demonstration of the parasite in tissue smears) or unable to distinguish between past and active infection (serological methods). Therefore, we aimed to develop a lateral flow assay in the form of an immunochromatographic test (ICT) device based on the detection of a circulating Leishmania antigen using monoclonal antibodies (mAbs).
  
  
  RESULTS
  mAbs were produced by fusion of murine myeloma cells with splenocytes isolated from a mouse immunized with L. donovani soluble crude antigen. Out of 12 cloned hybridoma cell lines, two secreted mAbs recognizing the same leishmanial protein. These mAbs were used to produce an ICT as a sandwich assay for the detection of circulating antigen in serum and blood samples. The ICT was evaluated with 213 serum samples from VL patients living in VL endemic areas in China, and with 156 serum samples from patients with other diseases as well as 78 serum samples from healthy donors. Sensitivity, specificity and diagnostic efficiency of the new ICT was 95.8%, 98.7% and 97.3%, respectively. Compared with a commercially available antibody detecting ICT, our antigen-based ICT performed slightly better.
  
  
  CONCLUSIONS
  The newly developed ICT is an easy to use and more accurate diagnostic tool which fulfils the performance and operational characteristics required for VL case detection under field and laboratory conditions. As our ICT detects a circulating antigen, it will also be useful in monitoring treatment success and diagnosing VL in immunocompromised patients.
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector, Ministry of Public Health, National Centre for International Research on Tropical Diseases, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, China.
authors.forename: Chun-hua
authors.initials: CH
authors.lastname: Gao
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector, Ministry of Public Health, National Centre for International Research on Tropical Diseases, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, China.
authors.forename: Yue-tao
authors.initials: YT
authors.lastname: Yang
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector, Ministry of Public Health, National Centre for International Research on Tropical Diseases, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Shi
authors.affiliation: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector, Ministry of Public Health, National Centre for International Research on Tropical Diseases, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, China.
authors.forename: Jun-yun
authors.initials: JY
authors.lastname: Wang
authors.affiliation: BioMedical Research Centre, Norwich Medical School, Norwich Research Park, University of East Anglia, Norwich, United Kingdom.
authors.forename: Dietmar
authors.initials: D
authors.lastname: Steverding
authors.affiliation: BioMedical Research Centre, Norwich Medical School, Norwich Research Park, University of East Anglia, Norwich, United Kingdom.
authors.forename: Xia
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology*; D000953:Antigens, Protozoan / Q000097:blood*; D002681:China; D002846:Chromatography, Affinity / Q000379:methods*; D005260:Female; D006801:Humans; D007893:Leishmania donovani / Q000276:immunology* / Q000302:isolation & purification; D007898:Leishmaniasis, Visceral / Q000175:diagnosis*; D051379:Mice; D008807:Mice, Inbred BALB C; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan