Colloidal Gold Immunochromatographic Assay for Rapid On-Site Detection of Tetracycline in Seawater

Recently increasing concerns from the scientists and public have been paid for seawater pollution due to tetracycline(TC)overuse in maricultural area.However,there are few methods or instruments that can be used for specific and rapid detection of this antibiotic in seawater.In this study,the colloidal gold immunochromatographic assay(CG-ICA)was used to achieve this goal.A commercialized monoclonal antibody against TC(anti-TC mAb)was selected because of its higher sensitivity(half-maximal inhibitory concentration of 2.38μgL^(-1)).The prepared CG particles(average diameter of 20 nm)were used to label anti-TC mAb at pH 8.0.The conjugate pad was formed by spraying the CG-labeled anti-TC mAb on a glass fibre membrane followed by proper dryness.The test pad was made by immobilizing artificial antigen and anti-mouse mAb in the test line and the control line,respectively,in a nitrocellulose membrane.The test strip,assembled with sample pad,conjugate pad,test pad and absorbent pad,could be used to detect TC during seawater sample flowing through these components in turn.The results could be observed by the naked eye in 10min.The visible limit of detection(vLOD)was 20μgL^(-1) for TC in seawater.The CG-ICA test results were in good agreement with those of liquid chromatography-tandem mass spectrometry(LC-MS/MS).The assay also showed that,oxytetracycline(OTC)and chlortetracycline(CTC),as the structural analogues of TC,did not interfere with TC determination.Furthermore,the TC concentration given by test strip could not be affected by the fluctuation of temperature(10℃–30℃),pH(7–9)and salinity(0–40)of seawater.Therefore,CG-ICA is a suitable tool for rapid,on-site,and semi-quantitative detection of TC in seawater.

Preparation of colloidal gold immunochromatography strip for detection of methamidophos residue

Methamidophos (Met) is a broad spectrum organophosphorus insecticide and acaricide.Even a trace of its residue is harmful to humans and many animals.In this study,the synthesis and identification of colloidal gold particles and antibody-colloidal gold conjugates were performed,and the preparation of colloidal gold immunochromatography strip was conducted for detection of Met residue.The size of colloidal gold particles was checked using a transmission electron microscope (TEM).The formation of antibody-coll...

Application of Colloidal Gold to Fluorophotometric Determination of Human IgG

A method for the determination of human immunoglobulin G（IgG） based on a colloidal gold label by fluorospectrophotometry was developed. The sandwich immunoreaction among goat-anti-human IgG, human IgG and goat-anti-human IgG labeled with colloidal gold nanoparticles was applied in this experiment. First, a sandwich im- munocomplex was formed on the surface of 96 well clear polystyrene high bind stripwellTM microplate. After the formation of the sandwich immunocomplex, a solution was added to dissociate the immunocomplex at room temperature. Then the solution of each well was transferred into the corresponding test tube. Thirdly, the rhodamine B solution was injected into each test tube. The rhodamine B chloraurate was extracted into ether that was measured with a spectrofluorometer. The experimental results indicate that the fluorescence intensity increased with the increase of human IgG concentration. The fluorescence intensity of rhodamine B chloraurate at 570 nm was proportional to the logarithm of human IgG concentration in a range from 10 to 5 × 10^5 ng/mL. It was shown that the determination of human IgG was easily made with the proposed fluorospectrophotometry.

Effects of 105 traditional Chinese medicines on the detection ofβ-agonists in medicine extracts and swine urine based on colloidal gold immunochromatographic assay

Colloidal gold immunochromatographic assay(CGIA)is commonly used for the on-site detection ofβ-agonists that are sometimes used illegally as feed additives in swine diets.However,few studies have evaluated the causes of false-positive results that sometimes occur when applying CGIA in agricultural settings.In this study,we investigated if this false-positive phenomenon is related to the addition of certain traditional Chinese medicines(TCMs)to swine feed.We established and verified an extraction method for TCMs,and then applied CGIA to detectβ-agonists in the extracts of 105 TCMs and in the urine of swine dosed with TCMs,respectively.Liquid chromatography-tandem mass spectrometry was used to validate the results of the urine samples tested positive forβ-agonists using CGIA.The results were also verified using TCMs and colloidal gold test strips produced by different manufacturers.The extracts of Citri Reticulatae Pericarpium Viride,Citri Reticulatae Pericarpium,Magnoliae Officinalis Cortex,Chaenomelis Fructus,and Rhodiolae Crenulatae Radix Et Rhizoma were tested positive forβ-agonists.Meanwhile,the addition of Citri Reticulatae Pericarpium Viride and Citri Reticulatae Pericarpium to swine feed resulted in false-positive results forβ-agonists in swine urine.The results provide a new way to explain false-positive CGIA results and provide valuable information for livestock feeding programs.

Interface Characteristics Between Colloidal Gold and Kaolinite Surface by XPS

The distribution of gold colloids in kaolinite and the interaction between gold and kaolinite surface were investigated by transmission electron mieroscotgy （TEM） and X-ray photoelectron spectroscopy （ XPS ）. There is strong interaction between the gold particles and the edge surfaces of kaolinite, in low pH solution, the edge surface of kaolinite is positively charged and electrostutic attrcactive force between colloide gold panicles and the positive edge surface of kaolinite woald facilitate the adsorption of colloidal gold particles onto the suface. TEM observation shows that the aggregate morphology of gold particles was dominated by particle-particle interaction and gold particles were adsorbed on the edge surface of kaolinite crystals , resulting from the electrostatic attractive force between colloidal gold particles and the positice surfaces of kaolinite. XPS data show that in Au4 f electron spectra there are four energy peaks related to gold, 83.8 eV, 85.7 eV, 87.5 eV, and 89.4 eV, respectively, which suggests that in chemical states there are metallic gold and Au bonded to O, similar to the form of Au2O3 , and composite Au2O3 is formed between the edge surface of kaolinite and colloidal gold surface.

A novel colloidal gold immunochromatography assay strip for the diagnosis of schistosomiasis japonica in domestic animals

Background:Schistosomiasis remains a major public health concern in China and an epidemiological survey has revealed that schistosome-infected bovines and goats are the main transmission sources for the disease.Therefore,development of a sensitive technique for the diagnosis of schistosomiasis in domestic animals is necessary.Method:A novel colloidal gold immunochromatography assay(GICA)strip was developed for detecting Schistosoma japonicum in domestic animals.The colloidal gold was conjugated with recombinant streptococcal protein G(rSPG).As the test and control lines,the schistosome soluble egg antigen and rSPG,respectively,were blotted on nitrocellulose membrane.Results:The lowest detectable serum dilution was 1∶640 for schistosome-infected buffaloes.The cross-reaction rate of GICA was 14.29%with Paramphistomum sp.in buffaloes,16.67%with Haemonchus sp.in goats,and 33.33%with Orientobilharzia sp.in goats.These results were slightly lower and similar to those obtained through ELISA.Moreover,the strips for detecting S.japonicum in mice,rabbits,buffaloes,and goats showed high sensitivity(100.00%,100.00%,100.00%,and 100.00%,respectively)and specificity(100.00%,100.00%,94.23%,and 88.64%,respectively).And the sensitivity or specificity of the GICA strips did not present any significant differences after storage for 12 months at room temperature.When compared with ELISA,the GICA strips exhibited similar sensitivity and specificity in the diagnosis of schistosomiasis in mice,rabbits,buffaloes,and goats.Besides,only 5μl of serum are required for the test and the detection can be completed within 5 min.Conclusion:This study is the first to develop a GICA strip using gold-rSPG conjugate for the diagnosing of schistosomiasis in domestic animals,and preliminary results showed that the developed strip may be suitable for large-scale screening of schistosomiasis in endemic areas.

A colloidal gold-based immunochromatographic strip for rapid detection of SARS-CoV-2 antibodies after vaccination

Vaccination interventions is consideredan important preventive measure to block the transmission of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)and protect the organism from pathogen infection effectively.However,a quick and accurate technique to evaluate the immune efficacy of the SARS-CoV-2 inactivated vaccine remains scarce.In this paper,an IgM-IgG antibody combined detection colloidal gold immunochromatography assay kit was optimized and developed,which can assess the efficacy of the inactivated SARS-CoV-2 vaccine.We collected fingertip blood samples from 3 vaccinees and 1 unvaccinated sample.The results showed that the proportion of antibody was high after the second shots immunization.The colloidal gold-based immunochromatographic strip is rapid,convenient and easy to operate.It can be used as an auxiliary method for preliminary evaluation of the antibody effect of vaccine recipients,and provide a reference index for the potential clinical application value of the vaccine.

Influence of Single Use and Combination of Reductants on the Size, Morphology and Growth Steps of Gold Nanoparticles in Colloidal Mixture

A comprehensive investigation on the formation mechanism of gold nanoparticles (AuNPs) in colloidal mixture obtained from the reduction of chloroauric acid (HAuCl4) solution using a single reducing agent (sodium citrate;process-I), (tannic acid;process-II), and a combination of two reducing agents (sodium citrate plus tannic acid;process-III) is reported. The growth steps at different time intervals during synthesis of colloidal AuNPs were monitored in situ and ex situ using various methods for all the three processes. The measurement of changes in the surface plasmon band position of colloidal AuNPs, along with dynamic light scattering results gave important information for the first assessing of particle size, shape and distribution. Besides, the size and morphological changes at different stages during different processes were also analyzed by transmission electron microscopy. The final Au particles of processes-I & II exhibited different shapes (spherical and nanowires) with particle size and nano wire diameter of 12 nm and 17 nm, respectively. Nevertheless, combination of two reductants (process-III) surprisingly leads to drastically reduced size (ca. 3 nm) with spherical morphology compared to their parent solutions with either of single reducing agent. This result clearly indicates that the combination of reductants has a significant influence on the particle size, morphology and formation mechanism.

Electrochemical Metalloimmunoassay Based on Enlargement of Gold Nanoparticle Label Treated with Ag Enhancement System

A novel sensitive electrochemical immunoassay with colloidal gold as the antibody labeling tag and subsequent signal amplification by silver enhancement is described. Colloidal gold was treated by a light-sensitive silver enhancement system which made silver deposit on the surface of colloidal gold(form Au/Ag core-shell structure), followed by the release of the metallic silver atoms anchored on the antibody by oxidative dissolution of them in an acidic solution and the indirect determination of the dissolved Ag + ions by anodic stripping voltammetry(ASV) at a carbon fiber microelectrode. The electrochemical signal is directly proportional to the amount of analyte(goat IgG) in the standard or a sample. The method was evaluated by means of a noncompetitive heterogeneous immunoassay of immunoglobulin G(IgG) with a concentration as low as 0.2 ng/mL. The high performance of the method is related to the sensitive ASV determination of silver(Ⅰ) at a carbon fiber microelectrode and to the release of a large number of Ag + ions from each silver shell anchored on the analyte(goat IgG).

Anion Adsorption on an Au Colloid Monolayer Based Cysteamine-Modified Gold Electrode

Anion adsorption behavior on Au colloid surface was investigated in virture of depositing monolayers of Au colloid on the self-assembled monolayers of cysteamine on a gold electrode. Po- tential-dependent anion adsorption-desorption waves via the nonfaradaic current were obtained by means of cyclic voltammetry at Au coltoid-modified gold electrodes in the potential range of -2 00-600 mV. The adsorption sequence in the order of adsorption peak potentials (Epa) is OH- >citrate3 ->H2PO4- >Cl->SO42->ClO4->NO3-. Among them, citrate3- exhibited an en- tirely irreversible adsorption. A rise in temperature can increase the rates of adsorption-desorp- tion and improve the reversibility of the adsorption-desorption of Cl-, SO42-, ClO4-, NO3- and H2PO-4. The adsorption peak potentials shifted more negatively for Ca. 63 mV as the anion con- centrations were increased by a decade factor. The change of pH from 7 to 1 slightly affected the adsorption peak potentials of Cl- and NO3-. Au colloids with a smaller size (16 nm) gave rise to a better reversibility of the adsorption-desorption process and lower adsorption currents. The ex- perimental results of citrate ions adsorption on Au colloid surface show that Au colloids with a smaller size prepared by sodium citrate method exhibited a higher stability in the solution in com- parison to those with larger sizes because of its higher ratio of charge/mass. In other words, the smaller gold nanoparticles are covered with citrate ions monolayer that can also be formed at larg- er gold nanoparticles by means of electrochemical scan.

PREPARATION OF POLYMER MICROSPHERES WITH PYRIDYL GROUP AND THEIR STABILIZED GOLD METALLIC COLLOIDS

Narrow disperse poly（ethyleneglycol dimethacrylate-co-4-vinylpyridine） （poly（EGDMA-co-4-VPy）） microspheres were prepared by distillation-precipitation copolymerization of ethyleneglycol dimethacrylate （EGDMA） and 4-vinylpyridine （4-VPy） with 2,2＇-azobisisobutyronitrile （AIBN） as initiator in neat acetonitrile. The polymer microspheres containing pyridyl group were then utilized as stabilizer for gold metallic colloids with the diameter around 7 nm, which were prepared by the in situ reduction of gold chloride trihydrate with sodium borohydride through the coordination of the pyridyl group on the gel layer and surface of the microsphere with the gold metallic nano-particles. The catalytic properties of the pyridyl- functionalized microsphere-stabilized gold metallic colloids and the behavior of the stabilized-catalyst for the recycling were investigated with reduction of 4-nitrophenol to 4-aminophenol as a model reaction.

Gold Nanoparticulate Thin Films Fabricated by the Electrostatic Self-Assembly Process

Gold colloids were prepared by citrate-induced reduction of hydrogen tetrachloroaurale, and gold nanoparticles were electrostatically self-assembled with poly( diallyldimethylammonium chloride) into multi-layer thin films on si/icon and quartz substrates. The paniculate thin films were characterized by UV-vis spea-troscopy, surface, enhanced Raman scattering, atomic force microscopy and resistivity measurements. Due to the interparticle coupling between individual gold particles, an obvious collective particle plasmon resonance was ob-served on UV-vis spectra , and the particulate thin films exhibited a strong SERS effect. For multilayer thin films with a high particle coverage on substrates , resistivity of the order of 10-4 Ω·cm was yielded.

Evaluation of Dynamic Changes of IgG and IgM in COVID-19 Patients by Different Detection Methods

Objective: To analyze the dynamic evaluation of chemiluminescence, colloidal gold, and immunofluorescence chromatography in detecting antibodies in COVID-19 patients within four weeks of infection, and to provide evidence for clinical application. Method: 74 patients with confirmed SARS-COV-2 infection in the local area were selected as the experimental group, while 231 patients with negative SARS-COV-2 results but not vaccinated with Covid19 vaccine were selected as the control group;during the first, second, third, and fourth weeks after enrollment in the experimental group, three methods were used to detect SARS-COV-2 IgG and IgM in patients’ blood: chemiluminescence method, colloidal gold antibody method, and immuno-fluorescence chromatography. In the control group, three methods were used to detect SARS-COV-2 IgG and IgM during physical examination for SARS-COV-2 nucleic acids. The ROC curve was drawn to analyze the value of each indicator in predicting SARS-COV-2 infection, and the kappa method was used to analyze the consistency of the detection results of each indicator. Results: There was no significant difference in the positive rates of SARS-COV-2 IgM and IgG antibodies detected by chemiluminescence, colloidal gold, and immunofluorescence chromatography during the four-week period (P > 0.05). The positive rates of SARS-COV-2 IgM and IgG antibodies detected by the three methods during the first week of infection were not higher than 60%;when the three methods were used to detect SARS-COV-2 IgM and IgG in vivo, the AUC diagnosed by the test results was less than 0.80 at the first week, the diagnostic efficacy of the three methods was above 0.95 from the second week to the fourth week, and the diagnostic efficacy of the three methods was higher than 0.97 at the fourth week. The diagnostic efficacy of the three methods was comparable;the three methods for detecting SARS-COV-2 IgM and IgG antibodies showed high consistency in four cycles. Conclusion: Chemiluminescence, colloidal gold, and immunofluorescence chromatography are highly consistent in the detection of SARS-COV-2 IgM and IgG antibodies, and can be used as an auxiliary diagnosis and efficacy observation of novel coronavirus infections according to the needs, but the positive rate of infected people in the first week is low.

Electrochemical Aptasensor Based on Prussian Blue-Chitosan-Glutaraldehyde for the Sensitive Determination of Tetracycline

In this paper, a novel and sensitive electrochemical aptasensor for detecting tetracycline(TET)with prussian blue(PB) as the label-free signal was fabricated. A PB-chitosan-glutaraldehyde(PB-CS-GA)system acting as the signal indicator was developed to improve the sensitivity of the electrochemical aptasensor.Firstly, the PB-CS-GA was fixed onto the glass carbon electrode surface. Then, colloidal gold nanoparticles(Au NPs) were droped onto the electrode to immobilize the anti-TET aptamer for preparation of the aptasensor.The stepwise assembly process of the aptasensor was characterized by cyclic voltammetry(C-V) and scanning electron microscope(SEM). The target TET captured onto the electrode induced the current response of the electrode due to the non-conducting biomoleculars. Under the optimum operating conditions, the response of differential pulse voltammetry(DPV) was used for detecting the concentration of TET. The proposed aptasensor showed a high sensitivity and a wide linear range of 109-～ 105-M and 105-～ 102-M with the correlation coefficients of 0.994 and 0.992, respectively. The detection limit was 3.2×1010-M(RSD 4.12%). Due to its rapidity, sensitivity and low cost, the proposed aptasensor could be used as a pre-scanning method in TET determination for the analysis of livestock products.

Scientific literature addressing detection of monosialoganglioside A 10-year bibliometric analysis

OBJECTIVE： The study was undertaken to explore a bibliometric approach to quantitatively assess the research on detection of monosialoganglioside from 2002 to 2011. DATA RETRIEVAL： A bibliometric analysis based on the publications on Web of Science was performed using key words such as ＂monosialoganglioside＂, ＂colloidal gold＂, ＂high performance liquid chromatography＂ and ＂detection＂. SELECTION CRITERIA： （1） Research articles on the detection of monosialoganglioside; （2） researches on human and animal fundamentals, clinical trials and case reports; （3） article types： article, review, proceedings paper, note, letter, editorial material, discussion, book chapter; （4） Publication year： 2002-2011. Exclusion criteria： （1） unrelated articles; （2） type of articles： correction; （3） articles from following databases： all databases related to social science and arts ＆ humanities in Web of Science were excluded. MAIN OUTCOME MEASURES： （1） distribution of subject areas; （2） number of publications annually; （3） document type and language of publications; （4） distribution of institutions; （5） distribution of output in journals; （6） the number of countries in which the article is published; （7） top cited paper. RESULTS： Overall population stands at 1 880 research articles addressing detection of monosialoganglioside in Web of Science during the study period. Articles （1 599） were the most frequently used document type comprising 85.05%, followed by meeting abstracts, reviews and proceedings papers. The distribution of subject categories showed that monosialoganglioside research covered both clinical and basic science research. The USA, Japan, and Italy were the three most productive countries, and the publication numbers in the USA were highest with 559 papers. The University of Milan, Nagoya University, and Kinki University are the most productive institutions regarding detection of monosialoganglioside. In 559 articles published by Americans, Medical College of Georgia ranked the first with 30 articles, followed by University of Medicine and Dentistry of New Jersey （28 articles）, Cornell University （24 articles） and Johns Hopkins University （24 articles）. In 442 articles published by Japanese, Nagoya University ranked the first with 40 articles, followed by Kinki University （36 articles）, and Dokkyo University （31 articles）. Though the total number of publications by Japanese is smaller than Americans, the top three institutions published more publications than American institutions. There is a markedly increase in the number of publications on the subject detection of monosialoganglioside in 2004, which the peak in the past 10 years. The valley bottom of the subject appeared in 2005. In total, the research is increased with time prolonged. Journal of Neurochemistry, Journal of Biological Chemistry and Journal of Neuroimmunology were core subject journals in monosialoganglioside studies. CONCLUSION： This study highlights the topics in detection of monosialoganglioside research that are being published around the world.

Characterization of colloidal arsenic at two abandoned gold mine sites in Nova Scotia, Canada, using asymmetric flow-field flow fractionation-inductively coupled plasma mass spectrometry

Asymmetric flow-field flow fractionation-inductively-coupled plasma-mass spectrometry was used to determine whether colloidal arsenic（As） exists in soil pore water and soil extract samples at two arsenic-contaminated abandoned gold mines（Montague and Goldenville, Nova Scotia）. Colloidal arsenic was found in 12 out of the 80 collected samples（= 15%）, and was primarily associated with iron（Fe） in the encountered colloids. The molar Fe/As ratios indicate that the colloids in some samples appeared to be discrete iron–arsenic minerals, whereas in other samples, they were more consistent with As-rich iron（oxy）hydroxides. Up to three discrete size fractions of colloidal As were encountered in the samples, with mean colloid diameters between 6 and 14 nm. The pore water samples only contained one size fraction of As-bearing colloids（around 6 nm diameter）, while larger As-bearing colloids were only encountered in soil extracts.

Study on the Vesiculation during Mouse Sperm Acrosome Reaction

The location Of the mono-membrane and the bi-membrane vesicles of mouse sperm wasidentified using Con A in conjugation with the colloidal gold. The observation showed that bothmono-membrane vesicles and outer layer of the bi-membrane vesicles come from the outeracrosome membrane.The inner membrane layer of the bi-member vesicles and residual membrane distributed among the vesicles are really foe plasmalemma.It is suggested that the outeracrosome membrane did not fuse with the plasmalemma during mouse sperm acrosome reaction and that both the mono-membrane and the bi-membrane vesicles of mouse sperm wereformed due to winding of the outer acrosome membrane.

A new preparation method of gold nanoparticles by intra-reduction of sodium gold(Ⅰ) sulfite

In this study, well-dispersed gold nanoparticles were prepared by using intra-molecular reduction of sodium gold sulfite, without using additional reductants and chloride free. The technical parameters including transformation temperature, pH, and concentration were optimized by the single-factor method as 90 C, pH 1, and 0.01 mmol/L [Na3Au（SO3）2], respectively. The resultant colloidal transmission electron microscopy images （TEM） and UV-vis absorption spectrophotometer spectra were acquired to check their properties, and the results show this kind of colloidal gold is controlled to 6 nm in sizes and has good stability in solution.

Rapid detection of fluoroquinolone residues in aquatic products based on a gold-labeled microwell immunochromatographic assay

Objectives:Fluoroquinolones(FQs)are widely used in aquaculture,and their residues have caused many problems threatening human health.Here,this study aims to develop a colloidal gold immunochromatographic strip based on gold-labeled microwells to screen the residues of FQs on site.Materials and methods:The Protein A Magarose Beads affinity chromatography method was adopted to purify the ascites against FQs.By using a strategy of heterologous coating antigen,different coating antigens are applied to detect FQs.The gold-labeled microwell immunochromatographic assay was used to improve the sensitivity of the test strip by the advanced reaction of antigen and antibody.Results:The antibodies were verified to be of high purity up to 99%,and the titer reached 1:1024000.The combination(enoxacin-OVA and the antibody)detected the 4 banned FQs(pefloxacin,PEF;norfloxacin,NOR;lomefloxacin,LOM;ofloxacin,OFL)with the 50%inhibiting concentration(IC50)values ranging from 1.3 to 2.1 ng/mL and cross-reactions ranging from 67.3%to 106.1%.The analysis of spiked crucian carp,silver carp,grass carp,and shrimp samples showed that the limit of detection for PEF,NOR,LOM,and OFL was 4μg/kg.A comparative study with liquid chromatography tandem mass spectrometry(LC-MS/MS)demonstrated that the assay provided an effective screening tool for the rapid detection of FQs residues.Conclusions:The results indicated that the test strip can realize full coverage recognition of the 4 banned FQs and has good accuracy,specifi-city,reproducibility,and stability;therefore,they are more suitable for rapid detection of FQs in aquatic products.

Gold modified microelectrode for direct tetracycline detection

The residues of tetracycline antibiotics in water have attracted many concerns due to their harmful impact to human health. This paper reports an electrochemical sensor for the determination of tetracycline (TC) by the microelectrode, which was fabricated by electrodeposited gold colloids on tungsten tip. Cyclic voltammerty was used to study the electrochemical behavior of TC on the microelectrode. Well anodic wave was obtained at about 1.5 V in acidic solutions. Electrochemical determination of tetracycline was investigated using microelectrode by cyclic voltammetry. Under optimized conditions, the calibration curves for TC were obtained. The oxidation peak currents were linearly related to TC concentrations in the range of 1–10 mg·L?1 and 10–100 mg·L?1, respectively. The detection limit was 0.09 mg·L?1 (S/N = 3).