Traditional Lateral Flow Immunoassay Research Articles

Colorimetric and Photothermal Dual-Modal Switching Lateral Flow Immunoassay Based on a Forced Dispersion Prussian Blue Nanocomposite for the Sensitive Detection of Prostate-Specific Antigen.

Prostate-specific antigen (PSA) is a key marker for a prostate cancer diagnosis. The low sensitivity of traditional lateral flow immunoassay (LFIA) methods makes them unsuitable for point-of-care testing. Herein, we designed a nanozyme by in situ growth of Prussian blue (PB) within the pores of dendritic mesoporous silica (DMSN). The PB was forcibly dispersed into the pores of DMSN, leading to an increase in exposed active sites. Consequently, the atom utilization is enhanced, resulting in superior peroxidase (POD)-like activity compared to that of cubic PB. Antibody-modified DMSN@PB nanozymes serve as immunological probes in an enzymatic-enhanced colorimetric and photothermal dual-signal LFIA for PSA detection. After systematic optimization, the LFIA based on DMSN@PB successfully achieves a 4-fold amplification of the colorimetric signal within 7 min through catalytic oxidation of the chromogenic substrate by POD-like activity. Moreover, DMSN@PB exhibits an excellent photothermal conversion ability under 808 nm laser irradiation. Accordingly, photothermal signals are introduced to improve the anti-interference ability and sensitivity of LFIA, exhibiting a wide linear range (1-40 ng mL-1) and a low PSA detection limit (0.202 ng mL-1), which satisfies the early detection level of prostate cancer. This research provides a more accurate and reliable visualization analysis methodology for the early diagnosis of prostate cancer.

Enhancing antibody detection sensitivity in lateral flow immunoassays using endospores of Bacillus subtilis as signal amplifiers

Antibody detection is the critical first step for tracking the spread of many diseases including COVID-19. Lateral flow immunoassay (LFIA) is the most commonly used method for rapid antibody detection because it is easy-to-use and inexpensive. However, LFIA has limited sensitivity when gold nanoparticles (AuNPs) are used as the signals. In this study, the endospores of Bacillus subtilis were used in combination with AuNP in a LFIA to detect antibodies. The endospores serve as a signal amplifier. The detection limit was about 10−8 M for anti-beta galactosidase antibody detection whereas the detection limit of conventional LFIA is about 10−6 M. Furthermore, the proposed methods have no additional user steps compared with the traditional LFIA. This method, therefore, improved the sensitivity 100-fold without compromising any advantages of LFIA. We believe that the proposed method will be useful for detection of antibodies against HIV, Zika virus, SARS-CoV-2, and so on.

Enhancing the stability and sensitivity of a lateral flow immunoassay using stabilized AuNPs for the simultaneous detection of sulfonamides and antibacterial synergists in chicken meat

Surface decoration of gold nanoparticles (AuNPs) effectively improves the stability and sensitivity of a traditional AuNP-based lateral flow immunoassay (LFIA). AuNPs decorated with citric acid, tannic acid, polydopamine, and polyethylene glycol (PEG) have been reported. However, these prepared nanoparticles have yet to be compared with each other in the same analytical system. Here, we synthesized stabilized AuNPs decorated with these four ligands and explored their properties to enhance the stability and sensitivity of LFIA. The results revealed that AuNPs@PEG was the optimal signal probe with an average AuNP diameter of 30 nm. It demonstrated significant advantages in tolerating extreme conditions (concentration of NaCl at 0.34 M) and improved sensitivity (twofold better than traditional LFIA). AuNPs@PEG-based LFIA showed cut-off values of 20–20,000 μg/kg for 18 sulfonamides and 1.6–200 μg/kg for antibacterial synergists in chicken meat, and the corresponding limits of detection were 2.0–548.2 μg/kg and 0.2–9.5 μg/kg. In conclusion, we are the first to investigate the properties of four stabilized AuNPs, and AuNPs@PEG shows good promise for use in LFIA.

Afterglow Nanoprobe-Enabled Quantitative Lateral Flow Immunoassay by a Palm-Size Device for Household Healthcare.

Lateral flow immunoassay (LFIA), a classical point-of-care testing (POCT) technique, plays an important role in disease screening and healthcare monitoring. However, traditional LFIA is either designed for qualitative analysis or requires expensive equipment for quantification, limiting its use in household diagnosis. In this study, we proposed a new generation of LFIA for household health monitoring by using ultralong organic phosphorescence (UOP) nanomaterials as afterglow nanoprobes with a self-developed palm-size sensing device. The UOP nanoprobes exhibit a phosphorescence signal with a second-level lifetime, which completely avoids the interference from excitation light and biological background fluorescence. Therefore, an ultraminiaturized and low-cost UOP nanosensor was successfully designed by eliminating the complex optical path and filtering systems. We chose an inflammatory factor, C-reactive protein (CRP), for household POCT validation. The whole analysis was completed within 9 min. A limit of detection (LOD) of 0.54 ng/mL of CRP antigen was achieved with high stability and good specificity, which is comparable to laboratory instruments and fully satisfying the clinical diagnosis requirement.

"Four-In-One" Multifunctional Dandelion-Like Gold@platinum Nanoparticles-Driven Multimodal Lateral Flow Immunoassay.

The traditional lateral flow immunoassay (LFIA) with a single signal output mode may encounter challenges such as low sensitivity, poor detection range, and susceptibility to external interferences. These limitations hinder its ability to meet the growing demand for advanced LFIA. To address these issues, the rational development of multifunctional labels for multimodal LFIA emerges as a promising strategy. Herein, this study reports a multimodal LFIA using "four-in-one" multifunctional dandelion-like gold@platinum nanoparticles (MDGP). The inherent properties of MDGP, such as the broad absorption spectrum, porous dandelion-like nanostructure, and bimetallic composition with gold and platinum, endow them with capacities in dual spectral-overlapped fluorescence quenching, optical readout, catalytic activity, and photothermal effect. Benefiting from their multifunctional properties, the MDGP-LFIA enables multimodal outputs including fluorescent, colorimetric, and photothermal signals. This multimodal MDGP-LFIA allows for the detection of acetamiprid at a range of 0.01-50ngmL-1 , with the lowest qualitative and quantitative detection results of 0.5 and 0.008ngmL-1 , respectively, significantly better than the traditional gold nanoparticles-based LFIA. The diversity, complementarity, and synergistic effect of integrated output signals in this multimodal MDGP-LFIA improve the flexibility, practicability, and accuracy of detection, holding great promise as a point-of-care testing platform in versatile application scenarios.

Molecular Engineering and Confinement Effect Powered Ultrabright Nanoparticles for Improving Sensitivity of Lateral Flow Immunoassay.

The application of traditional lateral flow immunoassay (LFIA)-based gold nanoparticles (AuNPs) to measure traces of target chemicals is usually challenging. In this study, we developed an integrated strategy based on molecular engineering and the spatial confinement of nanoparticles (NPs) to obtain ultrahigh quantum yields (QYs) of aggregation-induced emission (AIE) fluorescence NPs and employed them for the highly sensitive detection of T-2 toxin on the LFIA platform. Tetraethyl-4,4',4″,4‴-(ethene-1,1,2,2-tetrayl)tetrabenzoate (TCPEME), an AIE luminogen, was designed using molecular engineering to lower the energy gap, achieving higher QYs (26.26%) than previous AIEgens (13.02%). Subsequently, TCPEME-doped fluorescence NPs (TFNPs) achieved ultrahigh QYs, up to 84.55%, which were generated from the strong restriction of the NP state, efficiently suppressing nonradiative relaxation channels verified by ultrafast electron dynamics. On the LFIA platform, the sensitivity of the designed TFNP-based LFIA (TFNP-LFIA) was 10.4-fold and 4.3-fold more sensitive than that of the AuNP-LFIA and TPENP-LFIA for detecting the T-2 toxin, respectively. In addition, TFNP-LFIA was used for detecting T-2 toxin in samples and showed satisfactory recoveries (79.5 to 122.0%) with CV (1.49 to 11.75%), which implied excellent application potential for TFNP-LFIA. Overall, dual improvement of the molecule in fluorescence performance originating from the molecular engineering and spatial confinement of NPs could be an efficient tool for promoting the development of high-performance reporters in LFIA.

A Quantitative Detection Algorithm for Multi-Test Line Lateral Flow Immunoassay Applied in Smartphones.

The traditional lateral flow immunoassay (LFIA) detection method suffers from issues such as unstable detection results and low quantitative accuracy. In this study, we propose a novel multi-test line lateral flow immunoassay quantitative detection method using smartphone-based SAA immunoassay strips. Following the utilization of image processing techniques to extract and analyze the pigments on the immunoassay strips, quantitative analysis of the detection results was conducted. Experimental setups with controlled lighting conditions in a dark box were designed to capture samples using smartphones with different specifications for analysis. The algorithm's sensitivity and robustness were validated by introducing noise to the samples, and the detection performance on immunoassay strips using different algorithms was determined. The experimental results demonstrate that the proposed lateral flow immunoassay quantitative detection method based on image processing techniques achieves an accuracy rate of 94.23% on 260 samples, which is comparable to the traditional methods but with higher stability and lower algorithm complexity.

A universal boronate affinity capture-antibody-independent lateral flow immunoassay for point-of-care glycoprotein detection

Protein glycosylation and other post-translational modifications are involved in many biological processes including growth, development and immune responses, and glycoproteins are also known as biomarkers for cancer, diabetes and cardiovascular diseases. In traditional lateral flow immunoassay (LFIA) for glycoprotein detection, capture antibody (CA) is often required to label targets. However, the production of CA is complicated and expensive, restricting the wide application of LFIA. In this study, we developed a universal boronate affinity CA-independent LFIA method for glycoprotein detection. 4-Mercaptophenylboronic acid (4-MPBA)-modified Au nanoparticles (namely 4-MPBA-AuNPs) were used as LFIA labels, which could generate colorimetric signal and showed outstanding capability to bind glycoprotein. Compared with CA, 4-MPBA molecular as a glycoprotein recognition element had more prominent advantages, e.g., low cost, easy availability and good quality controllability. Take carcinoembryonic antigen (CEA) as model glycoprotein, the limit of detection of this CA-independent LFIA was 1.25 ng/mL by naked eyes, which was 8-fold lower than conventional CA-dependent sandwich LFIA. Significantly, the developed 4-MPBA-AuNPs-based CA-independent LFIA successfully detected 23 CEA-positive samples from 64 suspected human serum samples within 50 min in a nonlaboratory environment, with a 100% accuracy compared to clinical detection method. Therefore, this diagnostic platform could provide an effective tool for point-of-care glycoprotein detection with excellent reproducibility and high specificity.

Sharp Branched Gold Nanostar-Based Lateral-Flow Immunoassay for Detection of Yersinia pestis

Over the past few decades, colorimetric paper-based lateral flow immunoassay (LFIA) has emerged as a versatile analytical tool for rapid point-of-care detection of infectious diseases with high simplicity and flexibility. The LFIA sensitivity is based on color visualization of the antibody-labeled nanoparticles bound with the target analytes at the test line. Therefore, the nanoparticle design is crucial for LFIA sensitivity. The traditional LFIA is based on spherical gold nanoparticles, which usually suffer from poor sensitivity because of very low optical contrast at the test line. To improve the LFIA sensitivity, we have developed an LFIA based on gold nanostars (GNSs) with different branch lengths and sharpness (GNS-1, GNS-2, and GNS-3), which possess higher optical contrast than conventional gold nanospheres (GNSPs). We have selected the bacterium Yersinia pestis as a model analyte system. The effective affinity of GNSPs and GNSs with the Y. pestis fraction 1 (F1) protein was quantitively investigated by colorimetric and optical density measurements of the test line. The results show that GNS-3, which has maximum spike length and branch sharpness, exhibits the highest analytical sensitivity based on the limit of detection of the LFIA readout compared to other GNSs and GNSPs. The detection limit of the Y. pestis F1 antigen was achieved up to 0.1 ng/mL for GNS-3, which is 100 times lower than that for the GNSP at a 1 pmol/L concentration and 10 times lower than that for the reported procedure based on traditional gold nanoparticles. Overall, our prototype LFIA platform based on a highly spiked GNS (GNS-3) exhibits high analytical sensitivity, indicating it to be a promising candidate for routine LFIA application to detect infectious diseases.

Multimodal capture − antibody-independent lateral flow immunoassay based on AuNF − PMBA for point-of-care diagnosis of bacterial urinary tract infections

Bacterial urinary tract infections cause huge challenges to public health and require rapid diagnostic methods, while lateral flow immunoassay (LFIA) belongs to the most known point-of-care detection technology. Capture-antibody (CA) is used for labeling targets in traditional LFIA, yet the production of antibodies is time-consuming and expensive. In this study, we reported a multimodal CA-independent LFIA (MCI-LFIA) method for rapid diagnosis of bacterial urinary tract infections with high flexibility and accuracy. The assay employs p-mercaptophenylboronic acid-modified Au nanoflower (namely AuNF-PMBA nanomaterials) as multifunctional labels, which possess colorimetric-Raman-photothermal properties and outstanding bacteria capture ability. Take Escherichia coli (E. coli) as model bacteria, the limit of detection (LOD) of this MCI-LFIA for E. coli was 103 cfu/mL by naked-eye, which is three orders of magnitude lower than conventional CA-dependent sandwich method. For quantitative detection, LOD was 103 cfu/mL in colorimetric mode, 102 cfu/mL in Raman mode and 102 cfu/mL in photothermal mode. Significantly, the developed AuNF − PMBA-based MCI-LFIA successfully detected target pathogens in clinical E. coli-positive human urine samples within 45 min with high accuracy. This diagnostic platform may be easily adapted for detection of other target pathogens for bacterial diseases in point-of-care settings with good reproducibility and excellent specificity.

Detection of trh+ Vibrio parahaemolyticus in seafood by lac dye coloration-based label-free lateral flow immunoassay strip.

Vibrio parahaemolyticus (V.parahaemolyticus) is an important foodborne pathogen known to cause severe enteric disease. Thus, timely detection of V.parahaemolyticus in seafood is crucial to prevent food poisoning and reduce economic losses. Traditional lateral flow immunoassay strips (LFIS) required good labelling materials and pairing of two antibodies, which made them costly and difficult to manufacture. In this study, a label-free and lac dye coloration-based LFIS (LD-LFIS) to detect trh+ V.parahaemolyticus was developed for the first time. Lac dye was used to stain V.parahaemolyticus, and LFIS was used to detect stained bacteria. Dimethyl sulphoxide (DMSO) and simultaneous mordanting were chosen as the best solvent and the best staining method for lac dye. In addition, three mordants [KAl(SO4 )2 ·12H2 O, NH4 Fe(SO4 )2 ·12H2 O, and AlCl3 ·6H2 O] were selected to improve dyeing efficiency. The detection limit of LD-LFIS was 3.9 × 105 CFU/ml when NH4 Fe(SO4 )2 ·12H2 O was used as mordant. Feasibility of the LD-LFIS method was verified by detecting trh+ V.parahaemolyticus in true and spiked seafood samples. The method developed in this study is expected to reduce restrictions on labelling materials and pairing of two antibodies on LFIS, and proposes a novel idea for the rapid detection of V.parahaemolyticus in seafood.

Laser-patterned paper-based flow-through filters and lateral flow immunoassays to enable the detection of C-reactive protein

We report the use of a laser-based fabrication process in the creation of paper-based flow-through filters that when combined with a traditional lateral flow immunoassay provide an alternative pathway for the detection of a pre-determined analyte over a wide concentration range. The laser-patterned approach was used to create polymeric structures that alter the porosity of the paper to produce porous flow-through filters, with controllable levels of porosity. When located on the top of the front end of a lateral flow immunoassay the flow-through filters were shown to block particles (of known sizes of 200 nm, 500 nm, 1000 nm and 3000 nm) that exceed the effective pore size of the filter while allowing smaller particles to flow through onto a lateral flow immunoassay. The analyte detection is based on the use of a size-exclusive filter that retains a complex (∼3 μm in size) formed by the binding of the target analyte with two antibodies each of which is tagged with different-sized labels (40 nm Au-nanoparticles and 3 μm latex beads), and which is larger than the effective pore size of the filter. This method was tested for the detection of C-reactive protein in a broad concentration range from 10 ng/ml to 100,000 ng/ml with a limit-of-detection found at 13 ng/ml and unlike other reported methods used for analyte detection, with this technique we are able to counter the Hook effect which is a limiting factor in many lateral flow immunoassays.

Sensitive and hook effect–free lateral flow assay integrated with cascade signal transduction system

The traditional lateral flow immunoassay (LFI) for detecting foodborne pathogens suffers from low sensitivity and hook effect that led false–negative result. Here, we established a novel LFI integrated with a cascade signal transduction system for the sensitive and hook effect–free detection of Escherichia coli O157:H7 (E. coli O157:H7). First, E. coli O157:H7 was captured by immunomagnetic nanoparticles (IMNPs) and monoclonal antibody–β–lactamase–gold nanoparticles (ILAuNPs) forming sandwich complexes. Second, penicillin solution was added and hydrolyzed by β–lactamase on the sandwich complexes. Last, the penicillin hydrolysate was transferred to an ultrasensitive penicillin immunochromatographic test strip (PIST). With the high β–lactamase activity of ILAuNPs (0.1 μL of ILAuNPs hydrolyzing 20 ng/mL of penicillin) and ultrasensitive PIST (cutoff value achieving 2 ng/mL), the detection sensitivity of this method was 1.37 × 102 CFU/mL for E. coli O157:H7, which prevailed 1000 times over that of the traditional LFI. This work also efficiently solved hook effect when the E. coli O157:H7 concentration was over 107 CFU/mL. Thus, the detection range of the novel method was 102–108 CFU/mL, thereby expanding by four orders of magnitude than that of the traditional LFI (105–107 CFU/mL).

Integrated gold superparticles into lateral flow immunoassays for the rapid and sensitive detection of Escherichia coli O157:H7 in milk

Escherichia coli O157:H7 is a common harmful foodborne pathogen that can cause severe diseases at low infectious doses. Traditional lateral flow immunoassay (LFIA) for the rapid screening of E. coli O157:H7 in food suffers from low sensitivity due to its dependence on 20- to 40-nm gold nanoparticles (AuNP) with insufficient brightness as labels. To address this issue, we reported for the first time the successful synthesis of gold superparticles (GSP) by encapsulating numerous small AuNP into a polymer nanobead using an evaporation-induced self-assembly method. Results indicated that the resultant GSP exhibited remarkably enhanced absorbance compared with the most widely used 40 nm AuNP in LFIA. In addition, the absorbance of GSP could be easily tuned by varying GSP sizes. Under optimized conditions, we achieved a rapid and sensitive determination of E. coli O157:H7 in milk with a detection limit of 5.95 × 102 cfu/mL when using the GSP with a size of 342 nm as LFIA signal reporters, exhibiting improvement of approximately 32-fold relative to the conventional 40 nm AuNP-LFIA method. We further demonstrated the selectivity, accuracy, reliability, and practicality of the proposed GSP-LFIA strip. In summary, this work offers a new strategy for improving LFIA sensitivity using assembled GSP as markers and demonstrates huge potential in rapidly and sensitively detecting foodborne pathogens.

Self-assembled colloidal gold superparticles to enhance the sensitivity of lateral flow immunoassays with sandwich format.

Background: Traditional lateral flow immunoassay (LFIA) based on 20-40 nm gold nanoparticles (AuNPs) as signal reporter always suffers from relatively low detection sensitivity due to its insufficient brightness, severely restricting its wide-ranging application in the detection of target analytes with trace concentration.Methods: To address this problem, the self-assembled colloidal gold superparticles (GSPs) were synthesized as an improved absorption-dominated labeling probe for improving the sensitivity of sandwich LFIA. Five kinds of GSPs with the size ranging from 100 nm to 400 nm were synthesized by embedding hydrophobic AuNPs of size 12 nm as building blocks into the polymer nanobeads. The as-prepared GSPs were suggested as novel labeling probes of LFIA. The effects of the size of assembled GSPs on the sensitivity of sandwich LFIA was assessed, and the detection performance of GSPs-LFIA was further compared with traditional AuNPs-LFIA.Results: The resultant GSPs showed extremely high light absorption but very low light scattering, which favor the absorption-dominated signal output in LFIA. Among them, the GSP270-LFIA (size 270 nm) exhibits the highest sensitivity for human chorionic gonadotropin and hepatitis B surface antigen detection in real serum sample, which are approximate 39.79- and 13.8-fold higher than that of traditional AuNP40-LFIA.Conclusions: The proposed research demonstrated that the current GSPs can provide an ultrasensitive and quantitative detection for disease biomarkers in real serum samples as promising reporters of sandwich LFIA platform.

Ovalbumin antibody-based fluorometric immunochromatographic lateral flow assay using CdSe/ZnS quantum dot beads as label for determination ofT-2 toxin.

This work describes an anti-ovalbumin antibody-based lateral flow immunoassay (LFI) for T-2 toxin. The antibody uses acoating antigen as a bifunctional element for universality and introduces preincubation to improve the detection limits of the method. T-2 toxin and ovalbumin-modified T-2 toxin competitively binds on the anti-T-2 toxin monoclonal antibody modified on CdSe/ZnS quantum dot beads during preincubation. The modified T-2 toxin acts as a bifunctional element that forms immuno complexes during preincubation and combines with anti-ovalbumin antibody coated in the test line through the ovalbumin terminal. Fluorescence is detected at 610nm on the test zone following photoexcitation at 365nm. It has a reverse dose-effect relationship with the amount of T-2 toxin. The calibration plot is linear inthe 20-110fgmL-1T-2 toxin concentration range, and the limit of detection (LOD) is 10fgmL-1, which is lower by 8-fold than that of the traditional LFI system (LOD 80fgmL-1) and one order of magnitude than those of LFIs with labels of colloidal gold nanoparticles (LOD 150fgmL-1) or fluorophores (LOD 190ngmL-1). Universality was verified through aflatoxin B1 detection using the established ovalbumin antibody-based LFI system (LOD 10fgmL-1). The performance of the method was compared with that of established systems and a commercial ELISA kit (LOD 360fgmL-1). Graphical abstractSchematic representation of ovalbumin antibody-based immunochromatographic lateral flow assay for T-2 toxin. Preincubation is introduced for high sensitivity. T-2- anti-ovalbumin acts as a bi-functional element for universality. CdSe/ZnS quantum dot beads act as label. Fluorometric signal is detected at 610nm.

Invited review: Advancements in lateral flow immunoassays for screening hazardous substances in milk and milk powder

Dairy-related food safety outbreaks, such as food-borne pathogen contamination, mycotoxin contamination, and veterinary drug contamination, sometimes happen and have been reported all over the world, affecting human health and, in some cases, leading to death. Thus, rapid yet robust detection methods are needed to monitor milk and milk powder for the presence of hazardous substances. The lateral flow immunoassay (LFI) is widely used in onsite testing because of its rapidity, simplicity, and convenience. In this review, we describe some traditional LFI used to detect hazardous substances in milk and milk powder. Furthermore, we discuss recent advances in LFI that aim to improve sensitivity or detection efficiency. These advances include the use of novel label materials, development of signal amplification systems, design of multiplex detection systems, and the use of nucleic acid-based LFI.

Gold Nanoflowers and Gold Nanospheres as Labels in Lateral Flow Immunoassay of Procalcitonin

In this study, new types of nanogold labels for lateral flow immunoassay of model antigen procalcitonin based on photometric and surface-enhanced Raman scattering methods of detection were obtained. The linear range of procalcitonin determination applying gold nanoflowers as a label was between 0.5 and 10 ng/mL. The limit of photometric detection was achieved at 0.1 ng/mL that was five times lower than the sensitivity of traditional lateral flow immunoassay with spherical gold nanoparticles as a label. In addition, the conjugate of polyclonal antibodies against procalcitonin with spherical gold nanoparticles labeled with 4-mercaptobenzoic acid as Raman reporter molecule was prepared and used as an immunoprobe in lateral flow immunoassay based on surface-enhanced Raman spectroscopy to improve assay sensitivity.

Highly sensitive and selective lateral flow immunoassay based on magnetic nanoparticles for quantitative detection of carcinoembryonic antigen

Carcinoembryonic antigen (CEA) is an important biomarker in cancer diagnosis. Here, we present an efficient, selective lateral-flow immunoassay (LFIA) based on magnetic nanoparticles (MNPs) for in situ sensitive and accurate point-of-care detection of CEA. Signal amplification mechanism involved linking of detection MNPs with signal MNPs through biotin-modified single-stranded DNA (ssDNA) and streptavidin. To verify the effectiveness of this modified LFIA system, the sensitivity and specificity were evaluated. Sensitivity evaluation showed a broad detection range of 0.25–1000ng/ml for CEA protein by the modified LFIA, and the limit of detection (LOD) of the modified LFIA was 0.25ng/ml, thus producing significant increase in detection threshold compared with the traditional LFIA. The modified LFIA could selectively recognize CEA in presence of several interfering proteins. In addition, this newly developed assay was applied for quantitative detection of CEA in human serum specimens collected from 10 randomly selected patients. The modified LFIA system detected minimum 0.27ng/ml of CEA concentration in serum samples. The results were consistent with the clinical data obtained using commercial electrochemiluminescence immunoassay (ECLIA) (p<0.01). In conclusion, the MNPs based LFIA system not only demonstrated enhanced signal to noise ratio, it also detected CEA with higher sensitivity and selectivity, and thus has great potential to be commercially applied as a sensitive tumor marker filtration system.