Development of a colloidal gold based immunochromatographic strip for 8-OHdG detection / 生物工程学报

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.

Development of a colloidal gold-based immunochromatographic strip for rapid detection of Rice stripe virus / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Rice stripe virus (RSV) causes dramatic losses in rice production worldwide. In this study, two monoclonal antibodies (MAbs) 16E6 and 11C1 against RSV and a colloidal gold-based immunochromatographic strip were developed for specific, sensitive, and rapid detection of RSV in rice plant and planthopper samples. The MAb 16E6 was conjugated with colloidal gold and the MAb 11C1 was coated on the test line of the nitrocellulose membrane of the test strip. The specificity of the test strip was confirmed by a positive reaction to RSV-infected rice plants and small brown planthopper (SBPH), and negative reactions to five other rice viruses, healthy rice plants, four other vectors of five rice viruses, and non-viruliferous SBPH. Sensitivity analyses showed that the test strip could detect the virus in RSV-infected rice plant tissue crude extracts diluted to 1:20 480 (w/v, g/mL), and in individual viruliferous SBPH homogenate diluted to 1:2560 (individual SPBH/μL). The validity of the developed strip was further confirmed by tests using field-collected rice and SBPH samples. This newly developed test strip is a low-cost, fast, and easy-to-use tool for on-site detection of RSV infection during field epidemiological studies and paddy field surveys, and thus can benefit decision-making for RSV management in the field.

Comparative study on rapid dot-immunogold staining and two immunogold silver staining assays for diagnosing schistosomiasis japonica.

A fast, specific, sensitive, convenient, and economical rapid-dot-immunogold staining (R-Dot-IGS) assay was used to detect serum antibodies in patients infected with Schistosoma japonicum. The soluble egg antigen of Schistosoma japonicum was added onto microspore membrane. After pre-reacting and blocking, the serum to be detected and sheep anti-human IgG labeled with chloroauric acid were added sequentially. The assay took 15 minutes. For comparison, the dot-immunogold silver staining (Dot-IGSS) and rapid micro-volume Dot-IGSS (RM-Dot-IGSS) assay were also performed. The positive rate to detect the serum of schistosomiasis japonica by the R-Dot-IGS, Dot-IGSS and RM-Dot-IGSS assay was 98%, 98% and 100%, respectively. Samples from 50 healthy controls, 10 cases of clonorchiasis, and 10 cases of paragonimiasis showed negative reactions except for one case of clonorchiasis with RM-Dot-IGSS assay. Compared with Dot-IGSS and RM-Dot-IGSS, R-Dot-IGS assay has similar sensitivity and specificity, but the latter is quicker, simpler, and cheaper. Therefore, R-Dot-IGS is strongly recommended for rapid diagnosis of schistosomiasis japonica both in epidemiological study and in the clinic.