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Performance of coumarin-derived dendrimer-based fluorescence-linked immunosorbent assay (FLISA) to detect malaria antigen.
Malar J. 2014 Jul 10; 13:266.MJ

Abstract

BACKGROUND

Due 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.

METHODS

Serially 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.

RESULTS

As 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).

CONCLUSION

FLISA has a keen and high performance to detect malaria antigen, suggesting a potential assay as malaria immunodiagnostic.

Links

PMC Free PDF

Authors+Show Affiliations

Yeo SJ
No affiliation info available
Huong DT
No affiliation info available
Han JH
No affiliation info available
Kim JY
No affiliation info available
Lee WJ
No affiliation info available
Shin HJ
No affiliation info available
Han ET
Zoonosis Research Center, Department of Infection Biology, School of Medicine, Wonkwang University, Iksan, Jeonbuk 570-749, Republic of Korea. ethan@kangwon.ac.kr.
Park H
No affiliation info available

MeSH

Amino Acid SequenceAntibodies, MonoclonalAntibodies, ProtozoanAntigens, ProtozoanCoumarinsDendrimersEnzyme-Linked Immunosorbent AssayEpitopesErythrocytesFluorescent Antibody Technique, DirectFluorescent DyesFluorometryHumansImmunoenzyme TechniquesImmunosorbent TechniquesL-Lactate DehydrogenaseMalaria, FalciparumMalaria, VivaxMolecular Sequence DataParasitemiaPlasmodiumPlasmodium falciparumPlasmodium vivaxProtozoan ProteinsSensitivity and SpecificitySequence AlignmentSequence Homology, Amino Acid

Pub Type(s)

Comparative Study
Journal Article
Research Support, Non-U.S. Gov't
Validation Study

Language

eng

PubMed ID

25011624

Citation

Yeo, Seon-Ju, et al. "Performance of Coumarin-derived Dendrimer-based Fluorescence-linked Immunosorbent Assay (FLISA) to Detect Malaria Antigen." Malaria Journal, vol. 13, 2014, p. 266.
Yeo SJ, Huong DT, Han JH, et al. Performance of coumarin-derived dendrimer-based fluorescence-linked immunosorbent assay (FLISA) to detect malaria antigen. Malar J. 2014;13:266.
Yeo, S. J., Huong, D. T., Han, J. H., Kim, J. Y., Lee, W. J., Shin, H. J., Han, E. T., & Park, H. (2014). Performance of coumarin-derived dendrimer-based fluorescence-linked immunosorbent assay (FLISA) to detect malaria antigen. Malaria Journal, 13, 266. https://doi.org/10.1186/1475-2875-13-266
Yeo SJ, et al. Performance of Coumarin-derived Dendrimer-based Fluorescence-linked Immunosorbent Assay (FLISA) to Detect Malaria Antigen. Malar J. 2014 Jul 10;13:266. PubMed PMID: 25011624.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Performance of coumarin-derived dendrimer-based fluorescence-linked immunosorbent assay (FLISA) to detect malaria antigen. AU - Yeo,Seon-Ju, AU - Huong,Dinh Thi, AU - Han,Jin-Hee, AU - Kim,Jung-Yeon, AU - Lee,Won-Ja, AU - Shin,Ho-Joon, AU - Han,Eun-Taek, AU - Park,Hyun, Y1 - 2014/07/10/ PY - 2014/05/20/received PY - 2014/07/04/accepted PY - 2014/7/12/entrez PY - 2014/7/12/pubmed PY - 2015/10/2/medline SP - 266 EP - 266 JF - Malaria journal JO - Malar J VL - 13 N2 - BACKGROUND: Due 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. METHODS: Serially 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. RESULTS: As 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). CONCLUSION: FLISA has a keen and high performance to detect malaria antigen, suggesting a potential assay as malaria immunodiagnostic. SN - 1475-2875 UR - https://www.unboundmedicine.com/medline/citation/25011624/Performance_of_coumarin_derived_dendrimer_based_fluorescence_linked_immunosorbent_assay__FLISA__to_detect_malaria_antigen_ DB - PRIME DP - Unbound Medicine ER -