Performance of coumarin-derived dendrimer-based fluorescence-linked immunosorbent assay (FLISA) to detect malaria antigen

Malaria Journal

Methodology

Eun-Taek Han¹  Jin-Hee Han¹  Ho-Joon Shin²  Jung-Yeon Kim³  Won-Ja Lee³  Hyun Park⁴  Dinh Thi Huong⁴  Seon-Ju Yeo⁴ 

[1] Department of Medical Environmental Biology and Tropical Medicine, School of Medicine, Kangwon National University, Chuncheon, Gangwon-do, Republic of Korea;Department of Microbiology, Ajou University School of medicine, 443-721, Suwon, Republic of Korea;Division of Malaria and Parasitic Disease, Korea National Institute of Health, 363-951, Osong, Republic of Korea;Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, 570-749, Iksan, Jeonbuk, Republic of Korea;

关键词: Coumarin-derived dendrimer;    Fluorescence-linked immunosorbent assay (FLISA);    Lactate dehydrogenase (LDH);    ELISA;    Plasmodium falciparum;    Plasmodium vivax;   

DOI  :  10.1186/1475-2875-13-266

 received in 2014-05-20, accepted in 2014-07-04,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundDue to limitation of conventional malaria diagnostics, including microscopy, polymerase chain reaction (PCR), and enzyme-linked immunosorbent assay (ELISA), alternative accurate diagnostics have been demanded for improvement of sensitivity and specificity.MethodsSerially diluted Plasmodium LDH antigens, Plasmodium falciparum-infected human red blood cells (RBC) derived from in vitro culture or patient’s samples were used for evaluation of the performance of fluorescence-linked immunosorbent assay (FLISA). Microscopic examination was used to determine parasite density and the performance of FLISA was compared to ELISA. Finally, sensitivity and specificity of FLISA was determined by human specimens infected with P. falciparum, Plasmodium vivax, Toxoplasma gondii, and amoebae.ResultsAs a result of FLISA, the fluorescent intensity was highly correlated with antigen amount and FLISA was more sensitive than ELISA. FLISA detected at least 0.01 ng/ml of pLDH antigen, which showed 1,000-fold higher sensitivity than ELISA. In vitro-cultured P. falciparum was detected up to 20 parasite number/μL in FLISA but 5120 parasite number/μLin sandwich ELISA. In vitro P. falciparum-infected RBC number was highly correlated with fluorescent intensity (R2 = 0.979), showing that FLISA was reliable for detection of P. falciparum and available for quantification of parasite numbers. Furthermore, eighteen patient samples infected with P. falciparum (n = 9) and P. vivax (n = 9) showed 100% of sensitivity (18/18). FLISA showed 96.3% of specificity (26/27) because one sample of patient blood infected with T. gondii gave a false positive reactivity among healthy donors (n = 9), T. gondii-infected patients (n = 9), and amoeba-infected patients (n = 9).ConclusionFLISA has a keen and high performance to detect malaria antigen, suggesting a potential assay as malaria immunodiagnostic.

【 授权许可】

Unknown   
© Yeo et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.