Study On The Detection Of Chlamydia Psittaci, By PCR And Epidemiological Investigation Of Chlamydia Psittaci In Chongqing

Zoonosis caused by Chlamydia psittaci (Chlamyida psittaci) is increasingly serious. However, it is lack of accurate and rapid diagnostic means. There is an urgent need to establish accurate and rapid PCR and RT-QPCR detection method of Chlamydia psittaci. In this study, PCR and RT-PCR were used to establish accurate and rapid detection method of Chlamydia psittaci; the PCR detection kit of Chlamydia psittaci was also developed. Compared with the IHA method, epidemiological investigation of Chlamydia psittaci was performed using PCR detection method established in this work. The quantitative relation between infection and detection of Chlamydia psittaci was preliminarily determined. The main results of this work are as follows:①The PCR and RT-QPCR detection methods of Chlamydia psittaci were established. Specific primers were designed on the basis of the major out membrane protein (momp) gene(PSITTACI 457203)in Chlamydophila psittaci. Using cs1 DNA extracted from duckas a template, the PCR detection of Chlamydia psittaci method was established by optimizing the reaction system,sensitivity can be reached template of 2.5×10-5 ng /μl. The PCR detection kit of Chlamydia psittaci was also developed.②RT-QPCR detection methods of Chlamydia psittaci were established. Using the plasmid DNA as template, real-time quantitative PCR was performed in two types of reaction instrument. Established by the Real-time quantitative PCR detection method, in the template concentration of 1.78×102-1.78×108 copy/μl, the linear correlation coefficient is 0.998; melting curves have only the peak value of 79.1℃Tm; approved , and inter-assay coefficient of variation (CV%) were 1.30-4.59% and 5.72-9.87%③Using real-time quantitative PCR detection method, the quantitative analysis was performed in mice (Mus musculus) infected with Chlamydia psittaci. The mice were randomly divided into seven groups, each of which contains 12 mice. One group was used as control, while other groups took peritoneal injection with Cps, sampled and detected daily. Thereafter, real-time PCR was applied to quatitatively detect the changes of Chlamydia psittaci in mice.④400 samples of serum and 430 corresponding tissue samples were randomly collected from poultry fed in different environment in six districts of Chongqing. IHA detected on the positive sera of 157 points to 157 in the corresponding tissue samples for PCR testing, a total of 146 samples were positive, the samples were positive rate of 93.0%. The results showed that the PCR detection method of Chlamydia psittaci had high specificity and stability.