Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA)
Lateral flow immunoassay (LFIA) is the most widely used platform of the point-of-care (POC) detection. Since the most important factor in deciding the results of this technique is the visual quality of the test, it should be at its optimum. To address this issue, the present study examines optimisin...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-01-01
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| Series: | Journal of Experimental Nanoscience |
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| Online Access: | http://dx.doi.org/10.1080/17458080.2020.1775197 |
| _version_ | 1819203673501204480 |
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| author | M. Amini M. R. Pourmand R. Faridi-Majidi M. Heiat M. A. Mohammad Nezhady M. Safari F. Noorbakhsh H. Baharifar |
| author_facet | M. Amini M. R. Pourmand R. Faridi-Majidi M. Heiat M. A. Mohammad Nezhady M. Safari F. Noorbakhsh H. Baharifar |
| author_sort | M. Amini |
| collection | DOAJ |
| description | Lateral flow immunoassay (LFIA) is the most widely used platform of the point-of-care (POC) detection. Since the most important factor in deciding the results of this technique is the visual quality of the test, it should be at its optimum. To address this issue, the present study examines optimising critical parameters to improve the quality of LFIA performance. In this regard the penicillin-binding protein 2a (PBP2a) in methicillin-resistant staphylococcus aureus (MRSA) was selected as a bacterial model. The targeted parameters included a) size of gold nanoparticles (AuNP), b) antibody conjugation conditions with AuNP, c) antibody concentrations in the capturing zones, d) the effects of blocking substances and different chemical buffers for pre-treatment of the strips. Visual quality of different manufactured strips revealed that the diameter of 14 ± 2, PH = 9.5 was the optimum for improvement of MRSA detection quality. The optimum concentration of labelling antibody in conjugate pad and concentration of the capturing antibody in the test band were 40 µg/ml and 1.1 µg/μl in PBS buffer (pH 7.4) respectively. In conclusion by finding the critical factors affecting the detection limit of lateral flow and optimising them, the visual quality of LFIA results can be increased. |
| first_indexed | 2024-12-23T04:23:39Z |
| format | Article |
| id | doaj.art-858e5c45f30e4d15bdf9959eb79c5170 |
| institution | Directory Open Access Journal |
| issn | 1745-8080 1745-8099 |
| language | English |
| last_indexed | 2024-12-23T04:23:39Z |
| publishDate | 2020-01-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Journal of Experimental Nanoscience |
| spelling | doaj.art-858e5c45f30e4d15bdf9959eb79c51702022-12-21T18:00:11ZengTaylor & Francis GroupJournal of Experimental Nanoscience1745-80801745-80992020-01-0115126627910.1080/17458080.2020.17751971775197Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA)M. Amini0M. R. Pourmand1R. Faridi-Majidi2M. Heiat3M. A. Mohammad Nezhady4M. Safari5F. Noorbakhsh6H. Baharifar7Department of Pathobiology, School of Public Health, Tehran University of Medical SciencesDepartment of Pathobiology, School of Public Health, Tehran University of Medical SciencesDepartment of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences (TUMS)Baqiyatallah Research Center for Gastroenterology and Liver Disease, Baqiyatallah University of Medical SciencesDepartment de Medicine, University de MontrealDepartment of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences (TUMS)Department of Immunology, Faculty of Medicine, Tehran University of Medical SciencesDepartment of Medical Nanotechnology, Applied biophotonics research center, Science and Research Branch, Islamic Azad UniversityLateral flow immunoassay (LFIA) is the most widely used platform of the point-of-care (POC) detection. Since the most important factor in deciding the results of this technique is the visual quality of the test, it should be at its optimum. To address this issue, the present study examines optimising critical parameters to improve the quality of LFIA performance. In this regard the penicillin-binding protein 2a (PBP2a) in methicillin-resistant staphylococcus aureus (MRSA) was selected as a bacterial model. The targeted parameters included a) size of gold nanoparticles (AuNP), b) antibody conjugation conditions with AuNP, c) antibody concentrations in the capturing zones, d) the effects of blocking substances and different chemical buffers for pre-treatment of the strips. Visual quality of different manufactured strips revealed that the diameter of 14 ± 2, PH = 9.5 was the optimum for improvement of MRSA detection quality. The optimum concentration of labelling antibody in conjugate pad and concentration of the capturing antibody in the test band were 40 µg/ml and 1.1 µg/μl in PBS buffer (pH 7.4) respectively. In conclusion by finding the critical factors affecting the detection limit of lateral flow and optimising them, the visual quality of LFIA results can be increased.http://dx.doi.org/10.1080/17458080.2020.1775197lateral flow immunoassaylfiagold nanoparticles (aunp)rapid detectionpbp2amrsa |
| spellingShingle | M. Amini M. R. Pourmand R. Faridi-Majidi M. Heiat M. A. Mohammad Nezhady M. Safari F. Noorbakhsh H. Baharifar Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) Journal of Experimental Nanoscience lateral flow immunoassay lfia gold nanoparticles (aunp) rapid detection pbp2a mrsa |
| title | Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) |
| title_full | Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) |
| title_fullStr | Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) |
| title_full_unstemmed | Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) |
| title_short | Optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of PBP2a in methicillin-resistant Staphylococcus aureus (MRSA) |
| title_sort | optimising effective parameters to improve performance quality in lateral flow immunoassay for detection of pbp2a in methicillin resistant staphylococcus aureus mrsa |
| topic | lateral flow immunoassay lfia gold nanoparticles (aunp) rapid detection pbp2a mrsa |
| url | http://dx.doi.org/10.1080/17458080.2020.1775197 |
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