Rapid detection of human heart-type fatty acid-binding protein in human plasma and blood using a colloidal gold-based lateral flow immunoassay.
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ABSTRACT:
SUBMITTER: Ren YG
PROVIDER: S-EPMC8438696 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Artemisinin-based combination therapy (ACT) plays an indispensable role in malaria control and elimination. However, the circulation of counterfeit, substandard drugs has greatly threatened malaria elimination campaigns. Most methods for the analysis of artemisinin and its derivatives require expensive equipment and sophisticated instrumentation. A convenient, easy-to-use diagnostic device for rapid evaluation of the quality of artemisinin drugs at the point-of-care is still lacking. In this study a lateral flow dipstick immunoassay was developed for qualitative and semi-quantitative analysis of artesunate (ATS) and dihydroartemisinin (DHA) in anti-malarial drugs.This assay was based on a monoclonal antibody (mAb) raised against ATS. ATS-bovine serum albumin and goat anti-mouse IgG, used as the test capture reagent and the control capture reagent, were coated on the nitrocellulose membrane to form the test line and control line, respectively. The conjugate pad was saturated with the gold-labelled anti-ATS mAb.The indicator range of the dipsticks, defined as lowest concentration of the target analytes between which the test line was not visible, were 100-200 and 200-500 ng mL(-1) for ATS and DHA, respectively. No competitive inhibition was observed up to 5,000 ng mL(-1) of quinine, chloroquine diphosphate salt, primaquine phosphate, pyrimethamine, lumefantrine, amodiaquine, piperaquine tetraphosphate tetrahydrate or pyronaridine tetraphosphate. Semi-quantitative analysis of ATS and DHA in commercial drugs and raw drug materials with the dipsticks produced result agreeable with those determined by high performance liquid chromatography (HPLC). Storage test showed that the indicator range for artemisinins remained unchanged after a week at 37 °C and increased four-folds after six months of storage at 4 °C or ambient temperature.The new selected mAb 3D82G7 with high avidity and broad cross reactivity for artemisinins was used to develop and optimize a dipstick immunoassay for qualitative and semi-quantitative analysis of ATS and DHA in anti-malarial drugs. The semi-quantitative analysis of ATS and DHA in commercial drugs and raw drug materials, and the specificity test of the artemisinin-related drugs both proved the accurate performance of the developed dipsticks for semi-quantitation of ACT samples. The dipstick may be used as a point-of-care device for identifying substandard and counterfeit ATS- and DHA-containing anti-malarial drugs.
| S-EPMC4113134 | biostudies-literature
Project description:Rice false smut is a worldwide devastating rice disease infected by the fungal pathogen Villosiclava virens. Ustiloxin A (UA) and ustiloxin B (UB), cyclopeptide mycotoxins, were the major ustiloxins isolated from the rice false smut balls (FSBs) that formed in the pathogen-infected rice spikelets. Based on the specific monoclonal antibodies (mAbs) 2D3G5 and 1B5A10, respectively, against UA and UB, the lateral flow immunoassays (LFIAs) were developed, and the indicator ranges for UA and UB both were 50-100 ng/mL. The cross-reactivities of UB for UA LFIA, and UA for UB LFIA were 5% and 20%, respectively, which were consistent with the icELISA results reported previously. Even at 50,000 ng/mL, none of other commonly existent metabolites in rice samples caused noticeable inhibition. The LFIAs were used for determination of UA and UB contents in rice FSBs and rice grains, and the results were agreeable with those by HPLC and icELISA. There was no change in the sensitivity of either dipstick stored at 4 °C) after at least three months. The developed LFIA has specificity and sensitivity for detecting UA and UB as well as simplicity to use. It will be a potential point-of-care device for rapid evaluation of the rice samples contaminated by UA and UB.
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Project description:Colloidal gold lateral flow immunoassay strips (AuNPs-LFIS) have been widely applied as qualitative diagnostic tools for point-of-care tests (POCT). If strip readers were incorporated, their use could be extended to quantitative analysis. However, their cost and non-portability render commercial strip readers unavailable for use in either home testing, community or rural hospital diagnosis. This is particularly true for on-site testing. Here, a smartphone-based reader was designed and 3D-printed for quantitatively assess AuNPs-LFIS. The basic principle of the devise was relying on a smartphone's ambient light sensor (SPALS). This sensor was harnessed to measure the transmitted light intensities originating from the T-lines on the strips, the transmitted light intensities vary with concentration of AuNP on the T-lines. To validate this approach, our newly developed smartphone's ambient light sensor-based reader (SPALS-reader) was used to readout AuNPs-LFIS of three analytical targets: cadmium ion (Cd2+; limit of detection (LOD) was 0.16 ng/mL), clenbuterol (CL; LOD was 0.046 ng/mL), and porcine epidemic diarrhea virus (PEDV; LOD was 0.055 μg/mL). The result showed good consistency with the results of conventional image analysis approaches, indicating that the smartphone-based device is appropriate for use in AuNPs-LFIS readouts. Compared with the traditional analysis method, the developed AuNPs-LFIS reader is easier operated, lower cost and more portable, which provided an on-site quantitative analysis tool for AuNPs-LFIS and enhances the applied range of AuNPs-LFIS.
| S-EPMC7127147 | biostudies-literature
Project description:The authors describe a gold nanocage-based lateral flow strip biosensor (LFSB) for low-cost and sensitive detection of IgG. This protein was used as a model analyte to demonstrate the proof-of-concept. The method combines the unique optical properties of gold nanocages (GNCs) with highly efficient chromatographic separation. A sandwich-type of immunoreactions occurs on the GNC-based LFSB which has the attractive features of avoiding multiple incubation, separation, and washing steps. The captured GNCs on the purple test zone and control zone of the biosensor are producing characteristic purple bands, and this enables IgG even to be visually detected. Quantitatation was accomplished by reading the intensities of the bands with a portable strip reader. The LFSB fabrication and assay parameters were optimized. The biosensor displays a linear response in the 0.5 to 50 ng·mL-1 IgG concentration range, and it has a 15 min assay time. The detection limit is 0.1 ng·mL-1 of IgG, which is 2.5 times lower than that when using a gold nanoparticle-based LFSB. In our perception, this assay has a wide potential for the detection of other proteins and species for which respective antibodies are available.
| S-EPMC5703415 | biostudies-literature
Project description:Flavivirus detection in humans and mosquito reservoirs has been an important issue since it can cause a variety of illnesses and could represent a health problem in geographical zones where the vector is endemic. In this work, we designed and characterized a biosensor based on gold nanoparticles (AuNPs) and antibody 4G2 for the detection of dengue virus (DENV) in vitro, obtaining different conjugates (with different antibody concentrations). The AuNP-4G2 conjugates at concentrations of 1, 3, and 6 µg/mL presented an increase in the average hydrodynamic diameter compared to the naked AuNPs. Also, as part of the characterization, differences in the UV-Vis absorbance spectrum and electrophoretic migration were observed between the conjugated AuNPs (with BSA or antibody) and naked AuNPs. Additionally, we used this biosensor (AuNP-4G2 conjugate with 3 µg/mL antibody) in the assembly of a competitive lateral flow assay (LFA) for the development of an alternative test to detect the flavivirus envelope protein in isolated DENV samples as a future tool for dengue detection (and other flaviviruses) in the mosquito vector (Aedesaegypti) for the identification of epidemic risk regions. Functionality tests were performed using Dengue virus 2 isolated solution (TCID50/mL = 4.58 × 103) as a positive sample and PBS buffer as a negative control. The results showed that it is possible to detect Dengue virus in vitro with this gold nanoparticle-based lateral flow assay with an estimated detection limit of 5.12 × 102 PFU. We suggest that this biosensor could be used as an additional detection tool by coupling it to different point-of-care tests (POCT) for the easy detection of other flaviviruses.
| S-EPMC9313084 | biostudies-literature
Project description:Chlorantraniliprole (CAP) is a new type of diamide insecticide that is mainly used to control lepidopteran pests. However, it has been proven to be hazardous to nontarget organisms, and the effects of its residues need to be monitored. In this study, five hybridoma cell lines were developed that produced anti-CAP monoclonal antibodies (mAbs), of which the mAb originating from the cell line 5C5B9 showed the highest sensitivity and was used to develop a gold nanoparticle-based lateral flow immunoassay (AuNP-LFIA) for CAP. The visible limit of detection of the AuNP-LFIA was 1.25 ng/mL, and the detection results were obtained in less than 10 min. The AuNP-LFIA showed no cross-reactivity for CAP analogs, except for tetraniliprole (50%) and cyclaniliprole (5%). In the detection of spiked and blind samples, the accuracy and reliability of the AuNP-LFIA were confirmed by a comparison with spiked concentrations and verified by ultra-performance liquid chromatography-tandem mass spectrometry. Thus, this study provides the core reagents for establishing CAP immunoassays and a AuNP-LFIA for the detection of residual CAP.
| S-EPMC10814772 | biostudies-literature
Project description:The use of lateral flow immunoassays (LFIAs) for rapid on-site testing is restricted by their relatively high limit of detection (LoD). One possible way to decrease the LoD is to optimize nanoparticle properties that are used as labels. We compare two types of Au nanoparticles: usual quasispherical gold nanoparticles (C-GNPs), obtained by the Turkevich-Frens method, and superspherical gold nanoparticles (S-GNPs), obtained by a progressive overgrowth technique. Average diameters were 18.6-47.5 nm for C-GNPs and 20.2-90.4 nm for S-GNPs. Cardiomarker troponin I was considered as the target analyte. Adsorption and covalent conjugation with antibodies were tested for both GNP types. For C-GNPs, the minimal LoD was obtained with 33.7 nm nanoparticles, reaching 12.7 ng/mL for covalent immobilization and 9.9 ng/mL for adsorption. The average diameter of S-GNPs varied from 20.2 to 64.5 nm, which resulted in a decrease in LoD for an LFIA of troponin I from 3.4 to 1.2 ng/mL for covalent immobilization and from 2.9 to 2.0 ng/mL for adsorption. Thus, we obtained an 8-fold decrease in LoD (9.9 to 1.2 ng/mL) by using S-GNPs. This effect can be related to more effective antibody immobilization and improved S-GNP optical properties. The obtained results can improve LFIAs for various practically significant analytes.
| S-EPMC7348961 | biostudies-literature
Project description:Last year, the novel coronavirus disease (COVID-19) emerged in Wuhan, and it has rapidly spread to many other countries and regions. COVID-19 exhibits a strong human-to-human transmission infectivity and could cause acute respiratory diseases. Asymptomatic carriers are able to infect other healthy persons, and this poses a challenge for public health; the World Health Organization (WHO) has already announced COVID-19 as a global pandemic. Nucleic acid testing, considered as the current primary method for diagnosing COVID-19, might lead to false negatives and is difficult to be applied for every suspected patient because of the existence of asymptomatic carriers. Meanwhile, detecting specific antibodies in blood, such as the IgM antibody, against the SARS-CoV-2 virus is another choice for COVID-19 diagnosis, as it is widely accepted that IgM is an important indicator in the acute infection period. In this study, a colloidal gold nanoparticle-based lateral-flow (AuNP-LF) assay was developed to achieve rapid diagnosis and on-site detection of the IgM antibody against the SARS-CoV-2 virus through the indirect immunochromatography method. For preparing AuNP-LF strips, the SARS-CoV-2 nucleoprotein (SARS-CoV-2 NP) was coated on an analytical membrane for sample capture, and antihuman IgM was conjugated with AuNPs to form the detecting reporter. Optimization of AuNP-LF assay was carried out by altering the pH value and the amount of antihuman IgM. The performance of AuNP-LF assay was evaluated by testing serum samples of COVID-19 patients and normal humans. The results were compared with the real-time polymerase chain reaction. The sensitivity and specificity of AuNP-LF assay were determined to be 100 and 93.3%, respectively, and an almost perfect agreement was exhibited by Kappa statistics (κ coefficient = 0.872). AuNP-LF assay showed outstanding selectivity in the detection of IgM against the SARS-CoV-2 virus with no interference from other viruses such as severe fever with thrombocytopenia syndrome virus (SFTSV) and dengue virus (DFV). AuNP-LF assay was able to achieve results within 15 min and needed only 10-20 μL serum for each test. As a whole, in the light of its advantages such as excellent specificity and stability, easy operation, low cost, and being less time-consuming, AuNP-LF assay is a feasible method for the diagnosis of COVID-19 in primary hospitals and laboratories, especially in emergency situations in which numerous samples need to be tested on time.
| S-EPMC7241732 | biostudies-literature
Project description:Grapevine leafroll-associated virus 3 (GLRaV-3) is one of the main pathogens of grapes, causing a significant loss in yield and decrease in quality for this agricultural plant. For efficient widespread control of this infection, rapid and simple analytical techniques of on-site testing are requested as a complementary addition for the currently applied hybridization (PCR) and immunoenzyme (ELISA) approaches. The given paper presents development and approbation of the immunochromatographic assay (ICA) for rapid detection of GLRaV-3. The ICA realizes a sandwich immunoassay format with the obtaining complexes ((antibody immobilized on immunochromatographic membrane)⁻(virus in the sample)⁻(antibody immobilized on gold nanoparticles (GNP)) during sample flow along the membrane compounds of the test strip. Three preparations of GNPs were compared for detection of GLRaV-3 at different dilutions of virus-containing sample. The GNPs with maximal average diameters of 51.0 ± 7.9 nm provide GLRaV-3 detection for its maximal dilutions, being 4 times more than when using GNPs with a diameter of 28.3 ± 3.3 nm, and 8 times more than when using GNPs with a diameter of 18.5 ± 3.3 nm. Test strips have been manufactured using the largest GNPs conjugated with anti-GLRaV-3 antibodies at a ratio of 1070:1. When testing samples containing other grape wine viruses, the test strips have not demonstrated staining in the test zone, which confirms the ICA specificity. The approbation of the manufactured test strips indicated that when using ELISA as a reference method, the developed ICA is characterized by a sensitivity of 100% and a specificity of 92%. If PCR is considered as a reference method, then the sensitivity of ICA is 93% and the specificity is 92%. The proposed ICA can be implemented in one stage without the use of any additional reactants or devices. The testing results can be obtained in 10 min and detected visually. It provides significant improvement in GLRaV-3 detection, and the presented approach can be transferred for the development of test systems for other grape wine pathogens.
| S-EPMC6315891 | biostudies-literature
Project description:Background: In industrialised countries diphtheria is a rare but still life-threatening disease with a recent increase in cases due to migration and zoonotic aspects. Due to the rarity of the disease, laboratory diagnosis of diphtheria is often carried out in central reference laboratories and involves the use of sophisticated equipment and specially trained personnel. The result of the diphtheria agent detection can usually be obtained after 5-6 days or more. Authors suggest a Lateral Flow Immunoassay (LFIA)-based laboratory algorithm for the diagnosis of diphtheria, which may render less time in issuing a result and could promote the testing be performed in laboratories closer to the patient. Methods: LFIA for diphtheria toxin (DT) detection was designed using a pair of monoclonal antibodies to receptor-binding subunit B of the DT, and validated with 322 corynebacterial cultures as well as 360 simulated diphtheria specimens. Simulated diphtheria specimens were obtained by spiking of human pharyngeal samples with test strains of corynebacteria. The simulated specimens were plated on selective tellurite agar and after 18-24 hours of incubation, grey/black colonies characteristic of the diphtheria corynebacteria were examined for the DT using LFIA. Results: The diagnostic sensitivity of the LFIA for DT detection on bacterial cultures was 99.35%, and the specificity was 100%. Also, the LFIA was positive for all pharyngeal samples with toxigenic strains and negative for all samples with non-toxigenic strains. For setting LFIA, a 6-hour culture on Elek broth was used; thus, under routine conditions, the causative agent of diphtheria could be detected within two working days after plating of the clinical specimen on the tellurite medium of primary inoculation. Conclusions: The availability of such a simple and reliable methodology will speed up and increase the accuracy of diphtheria diagnosis globally.
| S-EPMC10445807 | biostudies-literature