A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA
This paper describes a rapid, high-throughput flow-through membrane immunoassay (FMIA) platform. A nitrocellulose membrane was spotted in an array format with multiple capture and control reagents for each sample detection area, and assay steps were carried out by sequential aspiration of sample and...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2013-04-01
|
| Series: | Diagnostics |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2075-4418/3/2/244 |
| _version_ | 1798040257585741824 |
|---|---|
| author | Gonzalo J. Domingo Paul Yager Christiane Dolecek Jeremy Farrar Buddha Basnyat Stephen G. Baker Sabina Dongol Amit Arjyal Jennifer L. Osborn Katherine G. McKenzie Mitra Singhal Sujatha Ramachandran Barry Lutz |
| author_facet | Gonzalo J. Domingo Paul Yager Christiane Dolecek Jeremy Farrar Buddha Basnyat Stephen G. Baker Sabina Dongol Amit Arjyal Jennifer L. Osborn Katherine G. McKenzie Mitra Singhal Sujatha Ramachandran Barry Lutz |
| author_sort | Gonzalo J. Domingo |
| collection | DOAJ |
| description | This paper describes a rapid, high-throughput flow-through membrane immunoassay (FMIA) platform. A nitrocellulose membrane was spotted in an array format with multiple capture and control reagents for each sample detection area, and assay steps were carried out by sequential aspiration of sample and reagents through each detection area using a 96-well vacuum manifold. The FMIA provides an alternate assay format with several advantages over ELISA. The high surface area of the membrane permits high label concentration using gold labels, and the small pores and vacuum control provide rapid diffusion to reduce total assay time to ~30 min. All reagents used in the FMIA are compatible with dry storage without refrigeration. The results appear as colored spots on the membrane that can be quantified using a flatbed scanner. We demonstrate the platform for detection of IgM specific to lipopolysaccharides (LPS) derived from Salmonella Typhi. The FMIA format provides analytical results comparable to ELISA in less time, provides integrated assay controls, and allows compensation for specimen-to-specimen variability in background, which is a particular challenge for IgM assays. |
| first_indexed | 2024-04-11T22:04:59Z |
| format | Article |
| id | doaj.art-3e9369d81c01428eb4fe106024f7af14 |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-04-11T22:04:59Z |
| publishDate | 2013-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-3e9369d81c01428eb4fe106024f7af142022-12-22T04:00:44ZengMDPI AGDiagnostics2075-44182013-04-013224426010.3390/diagnostics3020244A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISAGonzalo J. DomingoPaul YagerChristiane DolecekJeremy FarrarBuddha BasnyatStephen G. BakerSabina DongolAmit ArjyalJennifer L. OsbornKatherine G. McKenzieMitra SinghalSujatha RamachandranBarry LutzThis paper describes a rapid, high-throughput flow-through membrane immunoassay (FMIA) platform. A nitrocellulose membrane was spotted in an array format with multiple capture and control reagents for each sample detection area, and assay steps were carried out by sequential aspiration of sample and reagents through each detection area using a 96-well vacuum manifold. The FMIA provides an alternate assay format with several advantages over ELISA. The high surface area of the membrane permits high label concentration using gold labels, and the small pores and vacuum control provide rapid diffusion to reduce total assay time to ~30 min. All reagents used in the FMIA are compatible with dry storage without refrigeration. The results appear as colored spots on the membrane that can be quantified using a flatbed scanner. We demonstrate the platform for detection of IgM specific to lipopolysaccharides (LPS) derived from Salmonella Typhi. The FMIA format provides analytical results comparable to ELISA in less time, provides integrated assay controls, and allows compensation for specimen-to-specimen variability in background, which is a particular challenge for IgM assays.http://www.mdpi.com/2075-4418/3/2/244flow-through membrane immunoassay (FMIA)enzyme-linked immunosorbent assay (ELISA)multiplexindirect IgM assaySalmonella enterica serovar Typhityphoidserodiagnosislow resource setting |
| spellingShingle | Gonzalo J. Domingo Paul Yager Christiane Dolecek Jeremy Farrar Buddha Basnyat Stephen G. Baker Sabina Dongol Amit Arjyal Jennifer L. Osborn Katherine G. McKenzie Mitra Singhal Sujatha Ramachandran Barry Lutz A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA Diagnostics flow-through membrane immunoassay (FMIA) enzyme-linked immunosorbent assay (ELISA) multiplex indirect IgM assay Salmonella enterica serovar Typhi typhoid serodiagnosis low resource setting |
| title | A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA |
| title_full | A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA |
| title_fullStr | A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA |
| title_full_unstemmed | A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA |
| title_short | A Rapid, Multiplexed, High-Throughput Flow-Through Membrane Immunoassay: A Convenient Alternative to ELISA |
| title_sort | rapid multiplexed high throughput flow through membrane immunoassay a convenient alternative to elisa |
| topic | flow-through membrane immunoassay (FMIA) enzyme-linked immunosorbent assay (ELISA) multiplex indirect IgM assay Salmonella enterica serovar Typhi typhoid serodiagnosis low resource setting |
| url | http://www.mdpi.com/2075-4418/3/2/244 |
| work_keys_str_mv | AT gonzalojdomingo arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT paulyager arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT christianedolecek arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT jeremyfarrar arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT buddhabasnyat arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT stephengbaker arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT sabinadongol arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT amitarjyal arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT jenniferlosborn arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT katherinegmckenzie arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT mitrasinghal arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT sujatharamachandran arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT barrylutz arapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT gonzalojdomingo rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT paulyager rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT christianedolecek rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT jeremyfarrar rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT buddhabasnyat rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT stephengbaker rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT sabinadongol rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT amitarjyal rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT jenniferlosborn rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT katherinegmckenzie rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT mitrasinghal rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT sujatharamachandran rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa AT barrylutz rapidmultiplexedhighthroughputflowthroughmembraneimmunoassayaconvenientalternativetoelisa |