Colloidal Gold Immunochromatographic Test Strip Research Articles

Application of gold immunochromatographic assay strip combined with digital evaluation for early detection of Toxoplasma gondii infection in multiple species

BackgroundTimely diagnosis of Toxoplasma gondii infection is necessary to prevent and control toxoplasmosis transmission. The gold immunochromatographic assay (GICA) is a means of rapidly detecting pathogen in samples. GICA-based diagnostic methods have been developed to accurately detect pathogens with high sensitivity and specificity, and their application in T. gondii diagnosis is expected to yield good results.MethodsColloidal gold test strips were produced using T. gondii C-terminal truncated apical membrane antigen 1 (AMA1C). Colloidal gold-AMA1C and colloidal gold-murine protein conjugate were synthesized under optimal conditions. A nitrocellulose membrane was treated with AMA1C and goat anti-mouse antibody as the test line and control line, respectively. In total, 90 cat serum samples were tested using AMA1C-GICA and a commercial enzyme linked immunosorbent assay (ELISA) kit. The GICA results were digitally displayed using a portable colloidal gold immunochromatographic test strip analyzer (HMREADER). The sensitivity, specificity, and stability of AMA1C-GICA were assessed, and this was then used to examine clinical samples, including 203 human sera, 266 cat sera, and 81 dog sera.ResultsAMA1C-GICA had a detection threshold of 1:32 for T. gondii-positive serum. The GICA strips specifically detected T. gondii antibodies and exhibited no reactivity with Plasmodium vivax, Paragonimus kellicotti, Schistosoma japonicum, Clonorchis sinensis, and Schistosoma mansoni. Consequently, 15 (16.7%) positive samples were detected using the AMA1C-GICA and commercial ELISA kits for each of the assays. The receiver-operating characteristic curve showed that GICA had a relative sensitivity of 85.3% and specificity of 92%, with an area under the curve of 98%. After analyzing clinical samples using HMREADER, 1.2%–23.4% of these samples were found to be positive for T. gondii.ConclusionsThis study presents a novel assay that enables timely and efficient detection of serum antibodies against T. gondii, thereby allowing for its early clinical diagnosis. Furthermore, the integration of digital detection using HMREADER can enhance the implementation of GICA.Graphical

A rapid and reliable immunochromatographic strip for detecting paraquat poinsoning in domestic water and real human samples

Paraquat (PQ) is one of the most commonly used herbicides, but it has polluted the environment and threatened human health through extensive and improper usage. Here, a new naked-eye PQ immunochromatographic strip was developed to recognize PQ in domestic water and real human samples within 10 min based on a novel custom-designed anti-PQ antibody. The PQ test strip could recognize PQ at a concentration as low as 10 ng/ml, reaching the high-efficiency time-of-flight mass spectrometry detection level and identifying trace amounts of PQ in samples treated with a diquat (DQ) and PQ mixture. Notably, both the performance evaluation and clinical trial of the proposed PQ strips were validated in multiple hospitals and public health agencies. Taken together, our study firstly provide the clinical PQ-targeted colloidal gold immunochromatographic test strip designed both for environment water and human sample detection with multiple advantages, which are ready for environmental monitoring and clinical practice.

Colloidal gold immunochromatographic test strip for virus detection: a review

Colloidal gold immunochromatographic strip is a fast, sensitive and accurate solid-phase labeling detection technology, which has the advantages of low price, easy operation, rapid detection and high specificity, with the potential to qualitatively detect the relevant viruses in a short time with desired sensitivity and accuracy. It effectively addresses the disadvantages of long detection time, equipment inconvenience and professionalism requirement of the traditional detection methods used in the medical, veterinary, animal, plant virus detection, pesticide residue detection and other areas. Presently, the technology has been applied in the detection of bacterial diseases, viral diseases and prevention of extensive spread of infectious diseases, and has sufficient room for further development. This review summarizes the application of colloidal gold immunochromatography strip for biological virus detection, followed by prospecting future perspectives.

Ultrasensitive detection of four organic arsenic compounds at the same time using a five-link cardboard-based assay

In order to effectively control the excessive use of organic arsenic reagents in livestock and poultry products, there is an urgent need to develop a method for rapid detection of multiple organic arsenic reagents. In this study, two haptens were designed and derivatized around the structural formula of roxarsone, and a highly-sensitive group-selective mAb 3F2 was prepared, which can simultaneously detect roxarsone, 4-aminophenylarsonic acid, 2-aminophenylarsonic acid and phenylarsonic acid. We further developed a colloidal gold immunochromatographic test strip (ICS) and prepared a five-link card that can simultaneously detect four organic arsenics in chicken and pork samples. Its quantitative detection limits (LOQ) for the four compounds in chicken and pork samples were 0.06 and 0.32 ng/mL, 0.11 and 0.29 ng/mL, 0.34 and 0.99 ng/mL, and 0.88 and 1.5 ng/mL, respectively. This multi-ICS detection provides a powerful tool for the on-site detection and rapid screening of organic arsenic reagents in actual samples.

Selection of early pregnancy specific proteins and development a rapid immunochromatographic test strip in cows

The most of embryo losses occur before the day 16 after artificial insemination, but there is no low cost and easy operation that can detect pregnancy with high accuracy within three weeks post-insemination in cattle. In this study, blood samples were collected at day 18 of the estrous cycle, and days 18, 25 and 35 of pregnancy, and relative levels of interferon stimulated genes (ISGs), Toll-like receptor (TLRs), complement components, early pregnancy factor (EPF) and pregnancy-associated plasma protein A (PAPPA) proteins were analyzed through Western blot. In addition, a colloidal gold immunochromatographic test strip was developed using the selected antibody, and the test was used for early pregnancy diagnosis. The results showed that there were changes in relative levels of plasma ISGs, TLRs, complement components, EPF and PAPPA proteins during early pregnancy in cattle, and complement component 1q (C1q) could be used as an ideal marker to develop a colloidal gold immunochromatographic test strip for early pregnancy diagnosis. In addition, the accuracy of pregnancy diagnosis by this test strip was 91.67% (11/12) for pregnant cows and 80% (8/10) nonpregnant cows at day 18 after insemination. In conclusion, the changes in plasma ISGs, TLRs, complement components, EPF and PAPPA proteins may be related to the maternal systemic immune modulation during early pregnancy, and a colloidal gold immunochromatographic test strip was developed for early pregnancy diagnosis using C1q as the ideal marker in cows. However, this colloidal gold immunochromatographic test strip needs further studies to improve the accuracy.

Rapid, on-site quantitative determination of higenamine in functional food using a time-resolved fluorescence microsphere test strip

Higenamine (HIG), a benzyltetrahydroisoquinoline alkaloid found naturally in plants, is classified as an S3 Prohibited Substance in the 2020 World Anti-Doping Agency (WADA) report. To avoid problems such as doping violations in competitive events, it is necessary to develop rapid and sensitive detection methods. In this study, a highly-sensitive anti-HIG monoclonal antibody (mAb) was prepared and a time-resolved fluorescent microsphere immunochromatographic test strip (TRFM-ICTS) was established for the rapid quantitative detection of HIG in functional foods. Under optimized conditions, the TRFM-ICTS was compared with colloidal gold immunochromatographic test strip (CG-ICTS), and the half-maximal inhibitory concentration (IC50) of TRFM-ICTS was 1.37 ng/mL. The spiked recoveries ranged from 86.4% to 105.3%, which was consistent with the results of liquid chromatography-tandem mass spectrometry (LC-MS/MS) in the detection of real functional food. Therefore, TRFM-ICTS can be a candidate method for doping monitoring in functional foods and a powerful tool for HIG quantification.

An ultrasensitive colloidal gold immunosensor to simultaneously detect 12 beta (2)-adrenergic agonists

In this study, we first prepared a selective monoclonal antibody against 12 beta (2)-adrenergic agonists (Salbutamol, Clenbuterol, Brombuterol, Clenpenterol, Mabuterol, Carbuterol, Cimbuterol, Mapenterol, Pirbuterol, Terbutaline, Cimaterol, and Clenproperol). Then three haptens were designed and derived, among which, haptenS3 used the amino group of the salbutamol analog to derive a carboxyl group containing a spacer, which is unique to this study. The half-maximal inhibitory concentration (IC50) values were 0.35 ng/mL (Salbutamol), 0.42 ng/mL (Clenbuterol), 0.78 ng/mL (Brombuterol), 0.88 ng/mL (Clenpenterol), 1.34 ng/mL (Mabuterol), 1.38 ng/mL (Carbuterol), 1.71 ng/mL (Cimbuterol), 2.24 ng/mL (Mapenterol), 2.25 ng/mL (Pirbuterol), 2.27 ng/mL (Terbutaline), 3.49 ng/mL (Cimaterol), and 4.89 ng/mL (Clenproperol). We further developed a monoclonal antibody-based colloidal gold immunochromatographic test strip for screening and detecting 12 beta (2)-adrenergic agonists in swine urine and lamb samples. The immunochromatographic method developed in this study is a suitable tool for the on-site rapid detection and screening of beta (2)-adrenergic agonists in swine urine and lamb samples.

Development of a colloidal gold immunochromatographic test strip for the rapid detection of iprodione.

Iprodione is a dicarboximide fungicide that is widely used in agriculture around the world. A reliable and rapid detection method is needed for the on-site monitoring of iprodione residues in a variety of agricultural products. Herein, a colloidal gold immunochromatographic test strip was developed based on a selected coating antigen and a specific monoclonal antibody against iprodione. The particle size of colloidal gold, the preparation technique of the conjugate pad, the composition of the loading buffer, and the extraction solvent were comprehensively optimized for the test strip. A cut-off value of 0.9 mg kg-1 (50 ng mL-1) and a visual limit of detection of 0.09 mg kg-1 (5 ng mL-1) were achieved in a complex matrix of tobacco. No cross-reactivity was observed for iprodione metabolite and four other widely used pesticides during tobacco growth. Furthermore, the developed colloidal gold immunochromatographic test strip was applied to determine iprodione residues in tobacco samples, and the obtained results were in good agreement with those obtained by liquid chromatography tandem mass spectrometry. Additionally, the test strip was found to be stable afterlong-term storage at 37 °C for two months. The developed colloidal gold immunochromatographic test strip showed excellent accuracy, sensitivity, specificity, and stability, therefore, it is suitable for the rapid detection of iprodione residues in complex matrices.

Development of an immunochromatographic test strip for the rapid detection of aristolochic acid A in herbal medicinal materials.

Plants containing aristolochic acid and its derivatives are nephrotoxic, mutagenic, and carcinogenic to humans; chronic diet poisoning caused by the aristolochic acid is the cause of endemic (Balkan) nephropathy and related cancers. To develop a colloidal gold immunochromatographic test strip (ICS) based on the competitive format for the rapid detection of aristolochic acid A (AA-A) in herbal medicinal materials. For the ICS based on gold nanoparticles (AuNPs), the antigen [AA-A-bovine serum albumin (BSA)], and goat anti-mouse IgG were drawn on the nitrocellulose membrane as the test line (T line) and the control line (C line), respectively. Monoclonal antibody (MAb)-AuNP conjugates were sprayed onto the conjugate pad. The sensitivity of the ICS was 6 ng/mL, and the test was completed in 10 min. The analysis of AA-A in traditional Chinese medicine samples showed that the ICS results were in good agreement with those obtained by high-performance liquid chromatography methods. These results demonstrated that the ICS test could be used as a reliable, rapid, cost-effective, and convenient qualitative tool for on-site screening techniques to detect AA-A in herbal medicinal materials without any special instrumentation.

Colloidal gold immunochromatographic test strips for broad-spectrum detection of Salmonella

Outbreaks of Salmonella infections have always attracted world-wide attention in food industry. Therefore, it is particularly important to detect Salmonella. In this study, we aimed to develop a rapid, simple, sensitive, and broad-spectrum colloidal gold immunochromatographic test strip (ICTS) for detecting Salmonella. The prepared monoclonal antibody (mAb) 1B4 was labeled with colloidal gold and used as the capture antibody, as it can pair with itself to form a sandwich detection platform. The sensitivity of the test strip for Salmonella typhimurium was 4 × 105 CFU/mL with naked eye and the pathogen was detected in 5–15 min. The colloidal gold ICTS can be used to detect 18 stains of Salmonella and there was no cross-reaction with 14 other food-borne pathogens. Salmonella typhimurium, Salmonella paratyphi B, and Salmonella enterica in chicken were detected after 7 h, 7 h and 6 h of incubation, respectively, and the detection limit was as low as 1 CFU/mL. In short, this is important reports of colloidal gold ICTS for detecting so many stains of Salmonella, and this will be a huge improvement for detecting multiple Salmonella in food samples.

A smartphone-based rapid quantitative detection platform for lateral flow strip of human chorionic gonadotropin with optimized image algorithm

Colloidal gold immunochromatographic test strip has been widely used as a rapid, simple and low-cost correct detection technology. However, its detection is often qualitative or semi-quantitative, which limits its clinical application to some extent. Herein, a portable test strip quantitative detection device based on smartphone to detect human chorionic gonadotropin (HCG) is developed. In experiment, a colloidal gold HCG detection strip based on antigen antibody immune response is constructed, and the quantitative results of three different image processing methods on the same strip detection are compared, including the threshold processing algorithm based on location information, the RGB color component extraction algorithm and the grayscale projection value processing algorithm, the results show that the last algorithm can realize the best recognition of the region of interest of strip. The mobile phone application software (App) based on this design shows that the detection limit of constructed colloidal gold HCG strip is 3 ng/mL with a linear range of 6–300 ng/mL. The detection result of real urine sample is consistent with the spiked concentration (R2 = 0.988), indicating that the concentration of HCG can be accurately measured in urine with this method, presenting the potential for instant diagnosis.

Development of a Colloidal Gold-based Immunochromatographic Test Strip for Detection of Cetacean Myoglobin

This protocol describes the development of a colloidal gold immunochromatographic test strip based on the sandwich format that can be used to differentiate the myoglobin (Mb) of cetaceans from that of seals and other animals. The strip provides rapid and on-the-spot screening for cetacean meat, thereby restraining its illegal trade and consumption. Two monoclonal antibodies (mAbs) with reactivity toward the Mb of cetaceans were developed. The amino acid sequences of Mb antigenic reactive regions from various animals were analyzed in order to design two synthetic peptides (a general peptide and a specific peptide) and thereafter raise the mAbs (subclass IgG1). The mAbs were selected from hybridomas screened by indirect ELISA, western blot and dot blot. CGF5H9 was specific to the Mbs of rabbits, dogs, pigs, cows, goats, and cetaceans while it showed weak to no affinity to the Mbs of chickens, tuna and seals. CSF1H13 can bind seals and cetaceans with strong affinity but showed no affinity to other animals. Cetacean samples from four families (Balaenopteridae, Delphinidae, Phocoenidae and Kogiidae) were used in this study, and the results indicated that these two mAbs have broad binding ability to Mbs from different cetaceans. These mAbs were applied on a sandwich-type colloidal gold immunochromatographic test strip. CGF5H9, which recognizes many species, was colloid gold-labeled and used as the detection antibody. CSF1H13, which was coated on the test zone, detected the presence of cetacean and seal Mbs. Muscle samples from tuna, chicken, seal, five species of terrestrial mammals and 15 species of cetaceans were tested in triplicate. All cetacean samples showed positive results and all the other samples showed negative results.

Development of a Colloidal Gold-based Immunochromatographic Test Strip for Detection of Cetacean Myoglobin.

This protocol describes the development of a colloidal gold immunochromatographic test strip based on the sandwich format that can be used to differentiate the myoglobin (Mb) of cetaceans from that of seals and other animals. The strip provides rapid and on-the-spot screening for cetacean meat, thereby restraining its illegal trade and consumption. Two monoclonal antibodies (mAbs) with reactivity toward the Mb of cetaceans were developed. The amino acid sequences of Mb antigenic reactive regions from various animals were analyzed in order to design two synthetic peptides (a general peptide and a specific peptide) and thereafter raise the mAbs (subclass IgG1). The mAbs were selected from hybridomas screened by indirect ELISA, western blot and dot blot. CGF5H9 was specific to the Mbs of rabbits, dogs, pigs, cows, goats, and cetaceans while it showed weak to no affinity to the Mbs of chickens, tuna and seals. CSF1H13 can bind seals and cetaceans with strong affinity but showed no affinity to other animals. Cetacean samples from four families (Balaenopteridae, Delphinidae, Phocoenidae and Kogiidae) were used in this study, and the results indicated that these two mAbs have broad binding ability to Mbs from different cetaceans. These mAbs were applied on a sandwich-type colloidal gold immunochromatographic test strip. CGF5H9, which recognizes many species, was colloid gold-labeled and used as the detection antibody. CSF1H13, which was coated on the test zone, detected the presence of cetacean and seal Mbs. Muscle samples from tuna, chicken, seal, five species of terrestrial mammals and 15 species of cetaceans were tested in triplicate. All cetacean samples showed positive results and all the other samples showed negative results.

A Modified Quantum Dot-Based Dot Blot Assay for Rapid Detection of Fish Pathogen Vibrio anguillarum.

Vibrio anguillarum, a devastating pathogen causing vibriosis among marine fish, is prevailing in worldwide fishery industries and accounts for grievous economic losses. Therefore, a rapid on-site detection and diagnostic technique for this pathogen is in urgent need. In this study, two mouse monoclonal antibodies (MAbs) against V. anguillarum, 6B3-C5 and 8G3-B5, were generated by using hybridoma technology and their isotypes were characterized. MAb 6B3-C5 was chosen as the detector antibody and conjugated with quantum dots. Based on MAb 6B3- C5 labeled with quantum dots, a modified dot blot assay was developed for the on-site determination of V. anguillarum. It was found that the method had no cross-reactivity with other than V. anguillarum bacteria. The detection limit (LOD) for V. anguillarum was 1 × 10(3) CFU/ml in cultured bacterial suspension samples, which was a 100-fold higher sensitivity than the reported colloidal gold immunochromatographic test strip. When V. anguillarum was mixed with turbot tissue homogenates, the LOD was 1 × 10(3) CFU/ml, suggesting that tissue homogenates did not influence the detection capabilities. Preenrichment with the tissue homogenates for 12 h could raise the LOD up to 1 × 10(2) CFU/ml, confirming the reliability of the method.

Advantages of fluorescent microspheres compared with colloidal gold as a label in immunochromatographic lateral flow assays

Label selection is of vital importance for immunochromatographic assays. In this study, the fluorescent microsphere test strip and colloidal gold immunochromatographic test strip (FM-ICTS and CG-ICTS) were developed for the detection of Escherichia coli O157:H7 on the basis of the sandwich format. Two types of labels, namely, colloidal gold particles (CG) and carboxyl-modified fluorescent microspheres (FMs), were compared while coupling with anti-E. coli O157:H7 monoclonal antibody (mAb). The FM-ICTS and CG-ICTS were also compared. Results show that the coupling rate between FMs and mAb was higher than that between CG and mAb. Under optimum conditions, the sensitivity of FM-ICTS was eight times higher than that of CG-ICTS. Approximately 0.1μg of mAb was used in every FM-ICTS, whereas 0.4μg of mAb was used in every CG-ICTS. The coefficient of variation of FM-ICTS and CG-ICTS was 4.8% and 16.7%, respectively. The FM-ICTS and CG-ICTS can be stored at room temperature for 12 months and specific to five E. coli O157:H7 strains. Milk sample inoculated with E. coli O157:H7 were tested by the FM-ICTS and CG-ICTS. The FM-ICTS sensitivity was 104CFU/ml while the CG-ICTS sensitivity was 105CFU/ml. The sensitivity, consumption of antibodies, and coefficient of variation of FM-ICTS were better than those of CG-ICTS for the detection of E. coli O157:H7.

Development of a colloidal gold immunochromatographic test strip for detection of lymphocystis disease virus in fish

To develop a gold immunochromatographic test strip for on-site rapid detection of lymphocystis disease virus (LCDV). Monodispersional colloidal gold and gold-labelled anti-LCDV monoclonal antibody (McAb) 2D11 were prepared and characterized by UV-visible spectroscopy and transmission electron microscopy. Gold-labelled probe was used as the detection antibody, and goat anti-mouse IgG at the control line and anti-LCDV McAb 1A8 at the test line of the test strip served as the capture antibody. The positive results could be easily judged by the presence of a red test line with naked eye within 10min. The test strip, in good agreement with enzyme-linked immunosorbent assay and dot-blotting in sensitivity and LCDV detection, gave a detection limit of 1μgml(-1) of LCDV and was stable for 6months at room temperature and 12months at 4°C. The test strip was specific, simple and convenient for rapid detection of LCDV presenting good stability and reproducibility. This ready-to-use test strip allows on-site rapid detection of LCDV in fish without the requirement of specialized equipments and professional personnel, which could augment the practical application for diagnosis of LCDV even in disadvantage areas.

Detection of survivin in esophageal cancer patients' serum with colloidal gold immunochromatography

Objective To investigate the feasibility of screening for esophageal cancer with survivin colloidal gold immunochromatographic test strip. Methods The serum samples from 158 esophageal cancer patients and 146 healthy individuals were tested with survivin colloidal gold immunoohromatographic test strips. Results 20 nm-diameter colloidal gold solution was prepared and survivin polyclonal antibodies with different concentrations were labeled to the gold particles. It showed that the optimal concentration for labeling was 12 μg/ml. The prepared survivin colloidal gold immunochromatographic strip teat showed positive rates of survivin in esophageal cancer group and control group were 51.9 % (82/158) and 15.1% (22/146) respectively (χ2 = 45.7, P 0.05), gender groups (male: 50.4%, female: 58.1%, χ2= 0.59, P > 0.05) , primary site groups (upper segment: 56.3%, middle segment: 46. 1%, lower segment: 32. 1%, χ2 = 2.64, P > 0.05), differentiation groups (well-differentiation: 56.3%, moderate differentiation: 43.2%, poor-differentiation: 32.7%,χ2= 1.63, P >0.05)and lymph node metastasis groups (with metastasis:43.3%, without metastasis:43.4%, χ2 = 0.00, P > 0.05). The sensitivity, specificity and accuracy of survivin colloidal gold immunochromatographic test strip was 51.9% (82/158), 84.9% (124/146) and 67.8% (206/304) respectively. Conclusions Survivin colloidal gold immunochromatographic test strip is fast, simple, easy-to-read. It could be used as a valuable tool for screening of high-risk patients with esophageal cancer. Key words: Survivin; Test strip; Colloidal gold; Immanochromatography; Esophageal cancer