Colloidal Gold Detection Research Articles

Research status of colloidal gold detection technology in medicine

Research status of colloidal gold detection technology in medicine

Preparation of Lateral Flow PVDF Membrane via Combined Vapor- and Non-Solvent-Induced Phase Separation (V-NIPS).

A large pore size Poly(vinylidene fluoride) (PVDF) membrane was prepared by the V-NIPS method using PVDF/N, N-dimethylacetamide (DMAc)/Polyvinyl pyrrolidone (PVP)/Polyethylene glycol (PEG) system. Firstly, the effect of different additive ratios on the membrane morphology and pore size was studied, and it was found that when the PVP:PEG ratio was 8:2, PVDF membranes with a relatively large pore size tend to be formed; the pore size is about 7.5 µm. Then, the effects of different exposure time on the membrane morphology and pore size were investigated, and it was found that as the vapor temperature increased, the pores on the surface of the membrane first became slightly smaller and then increased. Finally, the effects of different vapor temperatures on the membrane properties were discussed. The results showed that the as-prepared membrane exhibited suitable capillary flow rate and similar performance compared with a commercially available membrane in colloidal gold tests. The likely cause is that the amount of negative charge is less and the capillary migration rate is too fast. This paper provides a reference for the preparation of PVDF colloidal gold detection membrane.

An Universal Colloidal Gold Detection Method of Vitellogenin as an Environmental Estrogen Biomarker

An Universal Colloidal Gold Detection Method of Vitellogenin as an Environmental Estrogen Biomarker

A Smartphone-Based Detection Method of Colloidal Gold Immunochromatographic Strip

The outbreak of the new coronavirus (SARS-CoV-2) infection has become a global public health crisis. Antigen detection strips (colloidal gold) can be widely used in novel coronavirus clinical screening and can even be extended to home self-testing, which provides a practical and effective way for people to obtain health status information away from the crowd. In this paper, a colloidal gold detection system without complex devices is proposed, which is based on smartphone usage along with a mobile-phone software embedded with normalization algorithms and a special designed background paper. The basic principle of the device relies on image processing. First, the data of the green channel of the image captured by a smartphone are selected to be processed. Second, the calibration curves are established using standard black and white card, and the calibration values under different detection environments are obtained by calibration curves. Finally, to verify the validity of the proposed method, various standard solutions with different concentrations are tested. Results show that this method can eliminate the influence of different environments on the test results, the test results in different detection environments have good stability and the variation coefficients are less than 5%. It fully proves that the detection system designed in this paper can detect the result of colloidal gold immunochromatographic strip in time, conveniently and accurately in different environments.

Expression and purification of recombinant SARS-CoV-2 nucleocapsid protein in inclusion bodies and its application in serological detection

The current standard for the diagnosis of COVID-19 is the nucleic acid test of SARS-CoV-2 RNA, however, virus antibody detection has the advantages of convenient sample collection, high throughout, and low cost. When combining detection with nucleic acid detection, antibody detection can effectively compensate for nucleic acid detection. Virus infection always induce high antibody titer against SARS-CoV-2 nucleocapsid protein (N protein), which can be used to detect COVID-19 at both infected and convalescent patients. In this study we reported the expression and purification of N protein in E.coli from inclusion bodies by a combination of two cation exchange chromatography, and the yield of N protein was around 50 mg/L fermentation broth with more than 90% purity. A corresponding colloidal gold detection kit prepared with our purified N protein was used to verify the efficiency and accuracy our N protein in antibody detection method. Of the 58 COVID-19 PCR positive patients' inactivated serum samples, 40 samples were IgM positive (69.0%), and 42 samples were IgG positive (72.4%), and all 95 COVID-19 negative patients’ inactivated serum samples were both IgM and IgG negative. Our results indicates that the refolded soluble N protein could be used for the preliminary detection of IgG and IgM antibodies against SARS-CoV- 2.

Detection of microamounts of novel coronavirus residues in environment by digital PCR

<p indent="0mm">2019 novel coronaviruses (2019-nCoV) swept the world and are still at a high incidence stage. Highly sensitive and accurate nucleic acid molecular diagnostic techniques and methods have become a hot spot of global concern. 2019-nCoV is a kind of RNA virus. At present, there are many methods to detect virus RNA, including real-time fluorescence quantitative PCR (RT-PCR), colloidal gold detection, chemiluminescence detection, and so on. Colloidal gold detection and chemiluminescence detection need to use monoclonal antibodies against 2019-nCoV, but there are some problems in the preparation of 2019-nCoV monoclonal antibodies, such as long cycle, complex preparation and easy pollution. So the above two methods are not easy to be established for the detection of 2019-nCoV. The superiority of PCR technology is widely used in auxiliary medical clinical diagnosis, customs inspection and quarantine, the development of new agriculture, national defense military defense, and basic research of bioscience, and so on. However, the quantification of the target DNA fragment in RT-PCR is still relatively quantitative. Because this technique mainly depends on CT value and standard curve to quantify the target DNA in the experiment, which affects the amplification efficiency of the whole system, and the sensitivity and accuracy of the reaction system are greatly affected. Because the 2019-nCoV virus is highly contagious, the detection methodology is required to be highly sensitive, which is in line with the characteristics of digital PCR (droplet digital PCR system, referred to as ddPCR). ddPCR has the advantages of absolute quantification, high specificity, high sensitivity and strong interference ability, which can better combine molecular biology and medicine closely, and has a great advantage in the detection of 2019-nCoV. In this study, based on droplet digital PCR detection technology, the detection reagent and method were established using the principle of nested PCR. The Saliva of 333 2019-nCoV patients and their environmental samples were detected and compared with real-time PCR (RT-PCR). The linear range of the droplet digital PCR method established in this study was between 25–5 copies/µL, and minimum detection limit was 0.5 copies/µL, which was 2–3 orders of magnitude higher than that of the commercial real-time fluorescent quantitative PCR detection technology. In 333 samples, 7 were positive in 197 environmental samples, and 9 were positive in 136 human samples. The detection accuracy was 100%, and the environmental micro-virus residues that could not be detected by RT-PCR were detected. The digital PCR detection method established in this study was highly specific, and the minimum detection limit was far lower than that of RT-PCR. It is of great significance for sensitive and accurate detection of 2019-nCoV infection, early infection and environmental microvirus.

Seroprevalence and asymptomatic carrier status of SARS-CoV-2 in Wuhan City and other places of China.

Wuhan City (WH) in China was the first place to report COVID-19 in the world and the outbreak of COVID-19 was controlled in March of 2020 in WH. It is unclear what percentage of people were infected with SARS-CoV-2 and what percentage of population is carriers of SARS-CoV-2 in WH. We retrospectively analyzed the SARS-CoV-2 IgG and IgM antibody positive rates in 63,107 healthy individuals from WH and other places of China using commercial colloidal gold detection kits from March 6 to May 3, 2020. Statistical approaches were utilized to explore the difference and correlation for the seropositive rate of IgG and IgM antibody on the basis of sex, age group, geographic region and detection date. The total IgG and IgM antibody positive rate of SARS-CoV-2 was 1.68% (186/11,086) in WH, 0.59% (226/38,171) in Hubei Province without Wuhan (HB), and 0.38% (53/13,850) in the nation except for Hubei Province (CN), respectively. The IgM positive rate was 0.46% (51/11,086) in WH, 0.13% (51/38,171) in HB, and 0.07% (10/13,850) in CN. The incidence of IgM positive rates in healthy individuals increased from March 6 to May 3, 2020 in WH. Female and older age had a higher probability of becoming infected than males (OR = 1.34; 95% CI: 1.08–1.65) or younger age (OR = 2.25; 95% CI: 1.06–4.78). The seroprevalence of SARS-CoV-2 was relatively low in WH and other places of China, but it is significantly high in WH than other places of China; a large amount of asymptomatic carriers of SARS-CoV-2 existed after elimination of clinical cases of COVID-19 in Wuhan City. Therefore, SARS-CoV-2 may exist in a population without clinical cases for a long period.

Establishment and clinical performance evaluation of 2019 novel coronavirus antibody colloidal gold detection method

Objective To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig) M and IgG antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance. Methods A total of 278 patients who were treated at Wuhan Hankou Hospital and the People's Hospital of Honghu from February 12, 2020 to February 20, 2020 were collected. According to the diagnostic criteria, 89 patients were confirmed with 2019-nCoV nucleic acid positive diagnosis, and 189 were 2019-nCoV nucleic acid-negative suspected patients. A total of 273 medical examiners from Nanfang Hospital, Southern Medical University from 2015 to 2018 were selected as controls. The serum samples of patients were collected. 2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors. Indirect IgM and IgG colloidal gold techniques were established by using recombinant N protein. 2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction (RT-PCR) was used as control. Serum specimens were tested for 2019-nCoV IgM and IgG. The specificity and sensitivity of colloidal gold assay were analyzed. Results The sensitivity and specificity of IgM detection reagents were 78.7% and 98.2%, respectively, those of IgG detection reagents were 73.0% and 99.3%, respectively, and those of IgM combined with IgG detection were 87.6% and 98.2%, respectively. For suspected patients with negative 2019-nCoV nucleic acid, the positive rates of IgM and IgG were 59.8% (113/189) and 52.9% (100/189), respectively, and the positive rate of IgM combined with IgG detection was 66.1% (125/189). Conclusion This reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR. Key words: Coronavirus infection; Novel coronavirus; Gold colloid; Antibodies; Clinical performance

Clinical application of fecal occult blood testing of hemoglobin and transferrin at the same time

Objective To discuss the clinical value when using o-aniline, colloidal gold detection paper hemoglobin (Hb),transferrin colloidal gold detection paper (Tf) at the same time for fecal occult blood test to diagnose diseases of the gastrointestinal disease bleeding. Methods On 100 cases of healthy people specimens, with o-aniline, hemoglobin of colloidal gold paper, transferrin colloidal gold diagnostic test paper fecal occult blood test. Results Of 100 healthy people o-aniline positive rate of 12%, hemoglobin positive rate of 2.0%,transferrin positive rate of 2.0%, while the detection of hemoglobin and transferrin in the stool, positive rate is2.0%:152 cases of the disease have gastrointestinal bleeding in patients with o-toluidine positive rate of 69.5%,hemoglobin positive rates of 73.0%, transferrin positive rate of 88.2 %, while the detection of hemoglobin and transferrin in the stool, positive rate is 91.2%. Conclusions Immunity assay has high sensitivity and specificity. But hemoglobin due to the role of intestinal bacteria and the intestinal mucus composition of the infection and then cause variability, it can cause a false negative response. So use hemoglobin and transferring to mensurate gastrointestinal bleeding at the same time may increase the positive rate. Key words: Fecal occult blood; Transferrin; Hemoglobin; O-aniline

Immuno-electron microscopy reveals that the excitotoxin quinolinate is associated with the plasma membrane in human peripheral blood monocytes/macrophages.

Quinolinate (QUIN), a tryptophan-derived excitotoxin, was localized ultrastructurally in human peripheral blood monocytes/macrophages (MO) by immuno-electron microscopy. A combined carbodiimide/glutaraldehyde/paraformaldehyde-based fixation procedure was developed for optimal retention of QUIN in the cell as well as minimal loss of ultrastructure; a silver-enhanced colloidal gold detection system was used for electron-microscopic analysis. Gold particles representing QUIN immunoreactivity were associated with the inner side of the plasma membrane in normal MO. The number of gold particles increased significantly when QUIN levels were elevated by treatment with its precursor kynurenine, but location of the gold particles remained essentially the same under this condition. Treatment with interferon-gamma increased the number of Golgi bodies, vacuoles and pseudopodia, reflecting the activated state of the cell. Significantly increased numbers of gold particles representing QUIN were detectable in approximately the same location as in the case of kynurenine treatment. Combined treatment with kynurenine and interferon-gamma maximally increased the number of gold particles at the periphery of the cell. The pseudopodia were intensely stained with gold particles, while they were not detectable in the inner part of the cytoplasm or in any other organelle even under this activated condition. The significance of the specific location of QUIN revealed in the present study and its relation to the release and subsequent actions of QUIN are discussed.