Development a stacking pad design for enhancing the sensitivity of lateral flow immunoassay

Tsai, Tsung‐Ting; Huang, Tse-Hao; Chen, Chung‐An; Ho, Natalie Yi-Ju; Chou, Yu‐Min

doi:10.1038/s41598-018-35694-9

Cited by 79 publications

(66 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…For example, Anfossi et al [ 45 ] reported that the LFIA for ochratoxin A detection using a silver system was 10-fold more sensitive than the conventional gold-based LFIA that otherwise used the same immuno-reagents. However, the silver reagent was relatively unstable and highly light sensitive [ 16 ]. Recently, Tsai et al (2018) [ 16 ] reported the use of a stacking pad (additional membrane between the conjugation pad and test pad) for enhancing the sensitivity of the LFIA for detection of protein A and C-reactive protein.…”

Section: Discussionmentioning

confidence: 99%

“…However, the silver reagent was relatively unstable and highly light sensitive [ 16 ]. Recently, Tsai et al (2018) [ 16 ] reported the use of a stacking pad (additional membrane between the conjugation pad and test pad) for enhancing the sensitivity of the LFIA for detection of protein A and C-reactive protein. The stacking pad helped to extend the binding interaction of antigen and antibody.…”

Section: Discussionmentioning

confidence: 99%

“…A lateral flow immunoassay (LFIA) is a diagnostic device based on immunological reaction (antigen–antibody interaction) and chromatography (capillary action) of a labeled analyte (gold nanoparticles) through multiple membranes, including sample, conjugate, detection and absorbent pads [ 16 , 17 ]. This method is a low-cost, simple, rapid and portable detection device popular in several fields, such as agriculture, food, environmental sciences and biomedicine [ 18 ].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Development of a Prototype Lateral Flow Immunoassay for Rapid Detection of Staphylococcal Protein A in Positive Blood Culture Samples

et al. 2020

View full text Add to dashboard Cite

Bloodstream infection (BSI) is a major cause of mortality in hospitalized patients worldwide. Staphylococcus aureus is one of the most common pathogens found in BSI. The conventional workflow is time consuming. Therefore, we developed a lateral flow immunoassay (LFIA) for rapid detection of S. aureus-protein A in positive blood culture samples. A total of 90 clinical isolates including 58 S. aureus and 32 non-S. aureus were spiked in simulated blood samples. The antigens were extracted by a simple boiling method and diluted before being tested using the developed LFIA strips. The results were readable by naked eye within 15 min. The sensitivity of the developed LFIA was 87.9% (51/58) and the specificity was 93.8% (30/32). When bacterial colonies were used in the test, the LFIA provided higher sensitivity and specificity (94.8% and 100%, respectively). The detection limit of the LFIA was 107 CFU/mL. Initial evaluation of the LFIA in 20 positive blood culture bottles from hospitals showed 95% agreement with the routine methods. The LFIA is a rapid, simple and highly sensitive method. No sophisticated equipment is required. It has potential for routine detection particularly in low resource settings, contributing an early diagnosis that facilitates effective treatment and reduces disease progression.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Development of a Prototype Lateral Flow Immunoassay for Rapid Detection of Staphylococcal Protein A in Positive Blood Culture Samples

et al. 2020

View full text Add to dashboard Cite

show abstract

“…In this study, we introduced a multi-target lateral flow immunoassay strip (msLFIA), which added an additional membrane between the conjugation pad and the test pad to the conventional AuNP-based LFIA format to enhance the detection sensitivity 39 , for measuring the protein levels of alpha-defensin and CRP in synovial fluid samples from patients undergoing revision arthroplasty for septic or aseptic failure. In the previous study 39 , synovial fluid sample was pre-treated with a 1:100 dilution in PBS buffer because the pure synovial fluid is viscous and difficult to flow through the strip completely; however, in this study, we performed a quick preincubation of mixing the pure synovial fluid sample with the AuNPs-labeled antibodies, so the protein levels in synovial fluid are specific for analyte without dilution. We aimed to develop a multiplex LFIA for rapid and high performance for PJI diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Development of a multiplex and sensitive lateral flow immunoassay for the diagnosis of periprosthetic joint infection

Tsai

Huang

et al. 2019

Sci Rep

Self Cite

View full text Add to dashboard Cite

The diagnosis of periprosthetic joint infection (PJI) remains a challenge. However, recent studies showed that synovial fluid biomarkers have demonstrated greater diagnostic accuracy than the currently used PJI diagnostic tests. In many diagnostic tests, combining several biomarkers into panels is critical for improving diagnostic efficiency, enhancing the diagnostic precision for specific diseases, and reducing cost. In this study, we prove that combining alpha-defensin and C-reactive protein (CRP) as biomarkers possesses the potential to provide accurate PJI diagnosis. To further verify the result, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI. A total of 10 synovial fluid samples were tested using the msLFIA, and the results showed that the combined measurements of synovial fluid alpha-defensin and CRP levels were consistent with those obtained from a commercial enzyme-linked immunosorbent assay kit. In addition, we developed a multi-target lateral flow immunoassay strip (msLFIA) with staking pad design to obtain on-site rapid response for clinical diagnosis of PJI, which the multi-target design is used to increase specificity and the stacking pad design is to enhance detection sensitivity. As a result, the turnaround time of the highly sensitive test can be limited from several hours to 20 min. We expect that the developed msLFIA possesses the potential for routine monitoring of PJI as a convenient, low-cost, rapid and easy to use detection device for PJI.

show abstract

“…Whilst these sophisticated platforms achieve improved performance, they do involve either high-cost biochemical reagents such as enzymes 15,16 , external equipment such as electrophoresis apparatus 17,21 , complex fabrication 14 or multiple-step operation 4,10 . Simplified and more cost-effective methods to improve the sensitivity of LFA are needed to be truly implemented for the rapid in-situ detection 25 .…”

mentioning

confidence: 99%

Improvement in Detection Limit for Lateral Flow Assay of Biomacromolecules by Test-Zone Pre-enrichment

Zhang

Liu

Wang

et al. 2020

Sci Rep

View full text Add to dashboard Cite

Lateral flow assay (LFA) is one of the most prevalent commercially available techniques for point-of-care tests due to its simplicity, celerity, low cost and robust operation. However, conventional colorimetric LFAs have inferior limits of detection (LODs) compared to sophisticated laboratory-based assays. Here, we report a simple strategy of test-zone pre-enrichment to improve the LoD of LfA by loading samples before the conjugate pad assembly. The developed method enables visual LODs of miR-210 mimic and human chorionic gonadotropin protein, to be improved by 10-100 fold compared with a conventional LfA setup without introducing any additional instrument and reagent except for phosphate running buffer, while no obvious difference occurred for Aflatoxin B1 (AFB1). It takes about 6-8 min to enrich every 50 μL of sample diluted with phosphate running buffer, therefore we can get visual results within 20 min. We identified a parameter by modeling the entire process, the concentration of probe-analyte conjugate at test zone when signaling unit being loaded, to be important for the improvement of visual limit of detection. In addition, the test-zone pre-enrichment did not impair the selectivity when miR-210 mimic was adopted as target. Integrated with other optimization, amplification and modification of LfAs, the developed test-zone pre-enrichment method can be applied to further improve LoD of LfAs. Lateral flow assay (LFA), a paper-based in-situ detection platform, has been widely used in diverse fields such as biomedicine, environmental health, quality control and food safety due to its rapid, inexpensive, portable measurements 1. Basing on affinity interactions such as hybridization of oligonucleotide or aptamer-target, antibody-antigen or biotin-streptavidin, the signal of the test zone on the LFA test strip changes with the concentration of a particular analyte of interest when a liquid sample is applied 2,3. There are two basic formats of LFA: sandwich format and competitive format. In addition, the competitive format could be designed in the following two ways: fixing the capture probe of target at test zone (competitive format I) or fixing the competitive analogue at test zone (competitive format II). Generally, there are two strategies to load signaling unit in the conventional LFA. The first one is the direct sampling method with drying signaling unit on the conjugate pad fixed between the nitrocellulose (NC) membrane and the sample pad before sampling. This strategy is the most widely used and easily handled one and the results usually show significant concentration dependence. However, the limit of detection (LOD) is usually poor, which is the main drawback of this conventional LFA. The other strategy is the pre-mixing/pre-incubation sampling method with mixing signaling unit with sample solution to load together onto the sample pad 4. Signaling unit is greatly diluted by sample solution in this strategy, resulting in a prolonged interaction between signaling unit and capture molecules at test zone and ...

show abstract

Development a stacking pad design for enhancing the sensitivity of lateral flow immunoassay

Cited by 79 publications

References 52 publications

Development of a Prototype Lateral Flow Immunoassay for Rapid Detection of Staphylococcal Protein A in Positive Blood Culture Samples

Development of a Prototype Lateral Flow Immunoassay for Rapid Detection of Staphylococcal Protein A in Positive Blood Culture Samples

Development of a multiplex and sensitive lateral flow immunoassay for the diagnosis of periprosthetic joint infection

Improvement in Detection Limit for Lateral Flow Assay of Biomacromolecules by Test-Zone Pre-enrichment

Contact Info

Product

Resources

About