Study On Genital Tract Infection Model Of Mice By Chlamydia Trachomatis Serotype E

Objective: To establish a murine model of genital tract infection with C. t serotype E through vaginal inoculation in order to lay a foundation for the research of C. t vaccine. At the same time, know about the infection characteristic of murine genital tract infection model caused by C. t serotype E which belongs to human biovar of C.t.Methods:1, We adopted some useful methods to enhance the infection rate of C. t for C. t culture. Then we purified C. t EB with high speed centrifugation and got the concentration of purified EB. 2, We chose female BALB/c mice with the age of 6-7 weeks as our experimental animals. 10 and 3 days prior to inoculation with C. t serotype E through vagina, we administered progesterone to all the experimental mice by intramuscular injection. C.t infection in genital tract of infected mice was confirmed by 4 kinds of laboratory methods at certain times after inoculation. These laboratory methods included PCR, immunofluorescence, C.t isolation and IgG antibody detection in the blood by ELISA. 3, We inspected the elimination course of C. t serotye E from genital tract of C. t-infected mice by C. t culture. Indirect ELISA was used for detecting the IgG antibody to C. t after C. t inoculation then we compared IgG titers to find out the trend in limited times.Results:1, We cultured C. t serotype E successfully and got a C. tEB concentration of 5. 8952X108IFU/mL 2^ Murine infection model was established successfully confirmed by 4 kinds of laboratory methods. Firstly, electrophoresis using PCR products showed that there was a special strip located at about 500bp of marker, according with the objective gene of 517bp. Secondly, immunof luorescense showed there was bright-green punctate fluorescence in the epithelia of mice vaginal tract. Thirdly, C. t isolation from vaginal tract of all the C. t-infected mice was positive, fourthly, IgG antibody to C. t had been in the blood on 14 days post-inoculation detected by ELISA. 3> Challenge dose of 30ML108IFU/mL gave a 100% infection rate of mice. Elimination course of C. t serotype E shedding from genital tract was 7-14 days. 4^ We explored the optimal working condition of ELISA by which we detected the C. t IgG antibody. We discovered that C. t IgG antibody had exsited on 14 days post-infection, but the titer was low. With the time of post-inoculation going on, antibody titer grew slowly. On 35 days post-infection, antibody peaked(P<0. 05).ConclusionsrK culture C. t serotype E successfully and get the concentration of purified C. t EB. 2> Murine model of genital tract infection with C. t serotype E had been established successfully. We worked out the optimal inoculation dose. 3> Elimination course of C. t shedding from vagina was short. That C. t serotpye E was not the nature pathogen of mice contributes it. Vaginal inoculation of mice with human biovars of C. trachomatis produced a mild infection of the lower genital tract, which was characterized by the shedding of fewer chlamydiae for a shorter duration. 4^ We discovered titer trend of IgG antibody to C. t after only a single vagina challenge of C. t serotype...