The Studies Of Test Strip For Rapid Detection Of Three Kinds Of Rodent-Borne Diseases By GICA

Plague, hemorrhagic fever with renal syndrome and leptospirosis are major diseases in China,they are also the largest rodent-borne disease hazards.They are serious harm to public health. Before the establishment of New China, the plague rampant in the popular nearly half of a century, and make unremitting efforts to continuously reduce the number of annual incidence, but did not completely control, they are remaining potential threat; fever with renal syndrome and the hook end The disease, epidemic disease during the peak number of people have more than one hundred thousand per year in recent years, the incidence is among the 37 kinds of notifiable infectious diseases in the top ten. With China's sustained economic development, international and domestic trade and personnel exchanges have become increasingly frequent, adding to the plague, hemorrhagic fever with renal syndrome, leptospirosis, rat-borne diseases such as high rates of state, from mortality to see, they are 25 species A, B infectious diseases in the overall statutory, are within the top 10, not only seriously jeopardize the lives and health of the people, but also for social stability, industrial and agricultural production, economic development, tourism development, etc., causing great impact of prevention and treatment of infectious diseases is a key state in which the plague of Infectious Disease Law in the first part.In view of rodent-borne diseases are continuing high incidence of key areas, in a planned manner pathogenic rodent monitoring and testing, for early detection of epidemics, timely control has played a positive role. But in the field, test kit to monitor the vast majority of imported reagents, several sets of reagents, repeat sampling, respectively, to detect operation is relatively cumbersome and costly.Traditional microbial detection of bacteria or virus needs to smear, culture, afterword by the number of hours or days testing, operation cumbersome and complex and can not operate simultaneously detect a variety of microorganisms. Radioimmunoassay (RIA), ELISA analysis (EIA), fluorescence analysis (FIA), chemical analysis of the immune luminescence (CIA) and other microbial detection compared to traditional methods should be simple and quick, but also need a few hours, complex operation, and operation Members demanding. Due to the colloidal gold test strip detection of microorganisms with the traditional bacterial or viral smear, culture test, compared well with the RIA, ELISA, fluorescence analysis, chemical luminescence and other immunological detection methods compared to simple (one step test that is able to complete), and fast (within minutes), etc, so in microbial detection has great potential for development, but the test strip detection of micro-organisms can only be detected on individual microorganisms, and colloidal gold strip with a simultaneous detection of multiple multi-material testing, a joint inspection of the significance of the detection of multiple targets has great value, has become an increasingly rapid detection of pathogenic microorganisms strip of research.GICA technology is a colloidal gold labeling, immunoassay and chromatography analysis and other methods combined with the solid organic marker immunoassay. Its principle is:to strip fiber materials for the solid phase chromatography, the sample solution by capillary action so that mobility in the chromatography strip, while the sample analytes and chromatographic materials for the affected analytes body (antigen or antibody) occurs with high specificity, high affinity immune response, immune complexes during chromatography were trapped in the chromatography material enriched in a certain region (detection zone), the use of visual markers can be (colloidal gold) and obtained direct experimental phenomena (color). The marker is across the detection of free zone, and the combination of markers automatically separated. This analysis technique is simple rapid and can be one single determination, no special equipment and suitable for rapid on-site monitoring.By GICA detection method can detect multiple pathogens prepared diversified colloidal gold test strip for the detection of a significant number of indicators of the Joint Inspection great value, has become an increasingly rapid detection of pathogenic microorganisms strip of research. So far no simultaneous rapid detection of foreign plague, epidemic hemorrhagic fever, leptospirosis, rat-borne diseases such as three kinds of colloidal gold strip published.In this study, Yersinia pestis (Yersinia Pestis), Hantaan virus (Hantavirus; HV), leptospirosis (Leptospira) antigen fragments were optimized by computer analysis of homologous sequences defined a new antigenic sites. We Successfully constructed a number of pathogens by the three antigenic sites of tandem gene expression vector pET-20b-rYHL, in the prokaryotic expression vector system for expression, purification and identification of its activity. The purified protein was immune rabbit polyclonal antibody, followed by the colloidal gold immunochromatographic technology (GICA), antibody detection of three pathogens.The results showed that the target gene expression in BL21, a higher rate, and the purified protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) shows the relative molecular mass consistent with the design of the protein bands. Western blot, ELISA test showed that the recombinant protein and polyclonal antibodies can better combination of colloidal gold paper that could better detect antibodies to the three pathogens.Successfully established synchronization, rapid detection of plague, epidemic hemorrhagic fever, leptospirosis, rodent-borne diseases, the three detection methods, developed with independent intellectual property rights of colloidal gold strip, could be detected in serum, plasma and whole blood samples, a drop of Blood, an experiment testing the three rodent-borne diseases, low cost, without special equipment, to simple, fast, sensitive, specific, accurate and objective. Overcome the single-species detection reagents and the limitations of imported reagents for on-site rapid testing and detection of epidemiological surveys on many occasions, has broad application prospects.