Study On The Distribution And Pathogenesis Of The Plague Bacteria In The Infected Mice By The Up-conversion Phosphor Technique

Yersinia pestis could mainly cause sepsis bubonic plague, pneumonic plague and plague. Mainly manifested as fever, chills, headache, fatigue, and accompanied by severe toxemia symptom, body aches and irritability lymph node enlargement, pneumonia, bleeding tendency. In nature, the plague is mainly spread and propagation between rodents and flea animal. In the history of a total of three outbreaks of plague, second of which lasting for hundreds of years, resulting in tens of millions of deaths. And because of its features of spreading fast, strongly infectious and high fatality rate, the detection of Yersinia pestis and its infection in vivo in organs of pathogenic distribution and association study is very necessary. The main research contents of experiment were as follows:Thirty 8-weeks aged female BALB/c mice were selected to carry out the infection of the Yersinia pestis by intraperitoneal, intranasal and intravenous injection. Ten groups of mice were fed every 8 hours respectively to observe their situation(mental state, appetite, back hair, etc.)and in each group randomly selected mouse to be killed to get its tissues such as heart, liver, spleen, lung. Grinding the tissues respectively to prepare the samples and mixing the Yersinia pestis F1 antibody labled up-converting phosphor particles with them. Detecting the mixed samples by the flow cytometry to observe the distribution of plague bacteria in different organs of plague mice and discuss the pathogenesis of the disease. Objective:Applying the Up-converting Phosphor particles(UCPs) as the maker to immune assay. Making full use of its unique up- conversion luminescence properties to mark the Yersinia pestis F1 antibody and using the flow cytometry to detect the distribution of each organ in the diseased mice rapidly and sensitively. Methods: A series of modification and activation on the surface of UCP particles were carried out including amino modification and aldehyde modification to make the UCPs coupled with Yersinia pestis F1 antibody. Evaluating the results of the distribution of the Yersinia pestis in different organs detecting by the flow cytometry and using immunochromatography to prove the results. Results:(1) Optimized the modified process of UCPs, and using the transmission electron microscopy(TEM) to determinate that after the series of surface modifications, the size of UCPs were uniform with great dispersity, which has laid the foundation for the biological marker.(2) Establishing the up-converting phosphor technology-based lateral flow assay to detect the Yersinia pestis;(3) Establishing the up-converting phosphor technology-based flow cytometry assay to detect and analysis the Yersinia pestis rapidly. Conclusions:To use the up-conveting phosphor technology-based flow cytometry to detect the Yersinia pestis rapidly and accurately with good sensitivity and specificity. Offereing a powerful detective method to prevent biological terrorist attacks and to cope with the emergencies.