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Single-step, paper-based concentration and detection of a malaria biomarker.
Anal Chim Acta. 2015 Jul 02; 882:83-9.AC

Abstract

The lateral-flow immunoassay (LFA) is an inexpensive and rapid paper-based assay that can potentially detect infectious disease biomarkers in resource-poor settings. Despite its many advantages that make it suitable for point-of-care diagnosis, LFA is limited by its inferior sensitivity relative to sophisticated laboratory-based assays. Our group previously introduced the use of a micellar aqueous two-phase system (ATPS), comprised of the nonionic Triton X-114 surfactant, to concentrate biomarkers in a sample and enhance their detection with LFA. However, achieving complete phase separation and target concentration using the Triton X-114 system required many hours, and the concentrated sample needed to be manually extracted and applied to LFA. Here, we successfully integrated the concentration and detection steps into a single step that occurs entirely within a portable paper-based diagnostic strip. In a novel approach, we applied the micellar ATPS to a 3-D paper design and effectively reduced the macroscopic phase separation time from 8 h to approximately 3 min. The 3-D design was integrated with LFA to simultaneously concentrate and detect Plasmodium lactate dehydrogenase (pLDH), a malaria biomarker, in both phosphate-buffered saline and fetal bovine serum within 20 min at room temperature. Compared to a conventional LFA setup with a pLDH detection limit of 10 ng μL(-1), our single-step diagnostic successfully detected pLDH at 1.0 ng μL(-1), demonstrating a 10-fold detection limit improvement and resulting in a sensitive and user-friendly assay that can be used at the point-of-care. The integration of a micellar ATPS and LFA represents a new platform that can improve and promote the use of paper-based diagnostic assays for malaria and other diseases within resource-poor settings.

Authors+Show Affiliations

Department of Bioengineering, University of California, 420 Westwood Plaza, 5121J Engineering V, P.O. Box 951600, Los Angeles, CA 90095-1600, USA.Department of Bioengineering, University of California, 420 Westwood Plaza, 5121J Engineering V, P.O. Box 951600, Los Angeles, CA 90095-1600, USA.Department of Bioengineering, University of California, 420 Westwood Plaza, 5121J Engineering V, P.O. Box 951600, Los Angeles, CA 90095-1600, USA.Department of Bioengineering, University of California, 420 Westwood Plaza, 5121J Engineering V, P.O. Box 951600, Los Angeles, CA 90095-1600, USA; Division of Advanced Prosthodontics & Weintraub Center for Reconstructive Biotechnology, UCLA School of Dentistry, Los Angeles, CA 90095-1600, USA.Department of Bioengineering, University of California, 420 Westwood Plaza, 5121J Engineering V, P.O. Box 951600, Los Angeles, CA 90095-1600, USA. Electronic address: kamei@seas.ucla.edu.

Pub Type(s)

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

Language

eng

PubMed ID

26043095

Citation

Pereira, David Y., et al. "Single-step, Paper-based Concentration and Detection of a Malaria Biomarker." Analytica Chimica Acta, vol. 882, 2015, pp. 83-9.
Pereira DY, Chiu RY, Zhang SC, et al. Single-step, paper-based concentration and detection of a malaria biomarker. Anal Chim Acta. 2015;882:83-9.
Pereira, D. Y., Chiu, R. Y., Zhang, S. C., Wu, B. M., & Kamei, D. T. (2015). Single-step, paper-based concentration and detection of a malaria biomarker. Analytica Chimica Acta, 882, 83-9. https://doi.org/10.1016/j.aca.2015.04.040
Pereira DY, et al. Single-step, Paper-based Concentration and Detection of a Malaria Biomarker. Anal Chim Acta. 2015 Jul 2;882:83-9. PubMed PMID: 26043095.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Single-step, paper-based concentration and detection of a malaria biomarker. AU - Pereira,David Y, AU - Chiu,Ricky Y T, AU - Zhang,Samantha C L, AU - Wu,Benjamin M, AU - Kamei,Daniel T, Y1 - 2015/04/21/ PY - 2015/01/27/received PY - 2015/04/15/revised PY - 2015/04/20/accepted PY - 2015/6/5/entrez PY - 2015/6/5/pubmed PY - 2016/3/2/medline KW - Diagnostic KW - Lateral-flow immunoassay KW - Malaria KW - Micellar aqueous two-phase system KW - Triton X-114 SP - 83 EP - 9 JF - Analytica chimica acta JO - Anal. Chim. Acta VL - 882 N2 - The lateral-flow immunoassay (LFA) is an inexpensive and rapid paper-based assay that can potentially detect infectious disease biomarkers in resource-poor settings. Despite its many advantages that make it suitable for point-of-care diagnosis, LFA is limited by its inferior sensitivity relative to sophisticated laboratory-based assays. Our group previously introduced the use of a micellar aqueous two-phase system (ATPS), comprised of the nonionic Triton X-114 surfactant, to concentrate biomarkers in a sample and enhance their detection with LFA. However, achieving complete phase separation and target concentration using the Triton X-114 system required many hours, and the concentrated sample needed to be manually extracted and applied to LFA. Here, we successfully integrated the concentration and detection steps into a single step that occurs entirely within a portable paper-based diagnostic strip. In a novel approach, we applied the micellar ATPS to a 3-D paper design and effectively reduced the macroscopic phase separation time from 8 h to approximately 3 min. The 3-D design was integrated with LFA to simultaneously concentrate and detect Plasmodium lactate dehydrogenase (pLDH), a malaria biomarker, in both phosphate-buffered saline and fetal bovine serum within 20 min at room temperature. Compared to a conventional LFA setup with a pLDH detection limit of 10 ng μL(-1), our single-step diagnostic successfully detected pLDH at 1.0 ng μL(-1), demonstrating a 10-fold detection limit improvement and resulting in a sensitive and user-friendly assay that can be used at the point-of-care. The integration of a micellar ATPS and LFA represents a new platform that can improve and promote the use of paper-based diagnostic assays for malaria and other diseases within resource-poor settings. SN - 1873-4324 UR - https://www.unboundmedicine.com/medline/citation/26043095/Single_step_paper_based_concentration_and_detection_of_a_malaria_biomarker_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2670(15)00550-4 DB - PRIME DP - Unbound Medicine ER -
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