| Hepatitis C is a global epidemic of infectious diseases, the prevalence rate of about3%. About180000000people are infected with HCV, and the height of the chronicity is the biggest characteristic ofthe disease. In general, acute HCV infection will display the sub clinical state, and lead to chronic liverdisease. Patients often delay treatment in the case of the unconscious, which leads to the occurrence ofhepatocellular carcinoma and liver cirrhosis. And there is no HCV effects of antiviral drugs and vaccines.Therefore, control of HCV infection situation is grim. While the early discovery, early diagnosis and earlytreatment are the source of infection. Prevention and control of HCV interrupt transmission of the mosteffective means to the diagnosis of HCV, which put forward higher requirements. At present, the HCVscreening for antibody detection in serum, the main methods include: enzyme linked immunosorbent assay,polymerase chain reaction, recombinant immunoblot etc. The colloidal gold immunochromatographymethod compared with those methods is simple operation, convenient carrying, rapid, specific. And itdoes’t need any special equipment and reagents to obtain test results, especially suitable for field detection.In this paper, based on the colloidal gold principle of the immune response, developed a kind ofcolloidal gold immunochromatography test strip for rapid diagnosis of anti HCV antibody. The colloidalgold technique uses trisodium citrate reduction method to preparation colloidal gold, and use labellingHCV recombinant antigen preparation gold on Glass Fiber, while another recombinant HCV antigen andHCV antibody were used as the test line and control line protein coated nitrocellulose membrane, used forthe detection of anti the HCV antibody in serum.For the preparation of the specificity, sensitivity, stability good note, in the course of the study, considering the influence of various factors on the strip quality. The preparation of colloidal goldlabeled water, recombinant antigen concentration, pH value, labeled antigen antibody concentration,different cellulose nitrate membrane, different humidity environment on coated effect, different sizes ofcolloidal gold sensitivity factors such as differences of exploration and inquiry, ultimately determine thecolloidal gold using ultra pure water system preparation of the particle size of about40nm, marker proteinconcentration was20μg/mL, using Sartorius Stedim CN140without the backing film in the humidity of theenvironment is40%-60%, the control line and test line are respectively coated HCV monoclonal antibody1mg/mL and HCV recombinant antigen1.5mg/mL. To establishment the method of HCV colloidal golddouble antigen sandwich.To test the specificity, sensitivity, stability and other aspects of the system, and compared with ELISA.The results show the immune colloidal gold test strip is specificity, sensitivity, good stability, andconvenient, fast. The positive rate and negative rate of the colloidal gold test strip are good compared withELISA. Terefore, it can be used to detect HCV infection in blood donors, and has certain clinical value inresearch. |
