Study On Detection, Biovars, Inhibition Test And TetM Of Ureaplasma Urealyticum

Objective 1 To determine better methods that extract UUDNA among three methods . To understand if the differences exist between culture method and PCR method. To investigate UU infection in definite populations in our region, and to explore the relationships between UU and disease of genito-urinary tract.2 To investigate the infection of UU biovars in definite populations. 3 To determine the drug-resistance of UU to ten antibacterial agents and to study the effects of six sorts of Chinese medical herbs and three sorts of complexed herbs on clinical UU isolates.4 To study the relationships between UU tetM and tetracycline-phenotype resistance and understand the distrubution of UU strains carrying tetM in different UU biovars and population.Methods 1 UUDNA was extracted by ddH2O boiling lysis , prolease K method and phenol-chloroform method respectively and the results were further analyzed.Culture method and PCR method were respectively used to detect UU and the results were analyzed.The infection of UU were detected in specific populations by culture method. 2 PCR assays were applied to distinguish UU biovars.3 UU antibacteria susceptibility tests were carried out by direct brothdish methods .UU inhibitory tests in vitro to Chinese crude drugs were carried out by microbroth dilution methods.The results were based on the minimum inhibitory concentration.4 The detections of UUtetM gene were performed by PCR assays.Results 1 There was no difference between prolease K method and phenol -chloroform method.Between ddH2O boiling lysis and phenol-chloroform method,the former sensitive rate was lower.The difference of UU results between culture method and PCR method was no statistically significant.UU infection rate in STD population in our region was 39.5%,the infection rate in male was higher than that in the female. The UU infection in the population with urinary tract symptom , urine WBC increase,high vagina PH was higher than that in the population with no urinary tract symptom,urine WBC normal,low vagina PH respectively. Additionally ,the infection of UU assay in female population at younger age and with worse vagina sanitation was higher than that in female population at elder age and with worse vagina sanitation respectively. The infection of UU biovar 2 was 84.76%,UU biovar 1 was 15.24% in UU postive population in our region.Based on the statistical comparison between two results, the susceptibility rates of Minomycin, Azithromycin, Doxycline, Joxamycin, evofloxacin, Clarithromycin were higher than that of Roxithromycin, Tetracycline, Ofloxacin, Erythromycin respectively. The multidrug resistant rate in 226 clinical UU isolates was 47.79%. Chinese crude drugs'minimal inhibitory concentrations to the clinical UU strains were listed as the following: Chinese gall : 0.313～1.25g/L, gardenia: 0.625～5.00 g/L,chebulae: 1.25～5.00g/L, cortex phellodendri1:1.25～5.00 g/L, paeoniae radix: 1.25～2.50g/L,Banlangeng:10.00～40.00g/L, Bazhengsan: 2.50～20.00 g/L, Wulinsan: 2.50～20.00g/L, Liuheji: 0.625～1.25g/L. 4 The associativity existed between UU tetM and tetracycline phenotype -resistance.The tetM-positive in UU was 39.02% totally. The tetM-positive in UU was no statistical difference between two UU biovars, but the rate of UU isolates carrying tetM in STD population was higher than that in normal population.Conclusions 1 The extractions of UU DNA should choose prolease K method and phenol-chloroform method, rather than using ddH2O boiling lysis. 2 Culture method and PCR method were applied to UU assays, the results were coincident. 3 UU infection rate in STD population in our region was 39.5%, the infection rate in male was higher than that in the female.The populations with urinary tract symptom, urine WBC increase (with no urinary tract symptom), the females with high vagina PH, young females were high-risk UU infection groups. UU infection rate in STD population in our region was 39.5%,the infection rate in male was higher than that in female. the associativity existed between between UU infection and vagina sanitation grades. The probablity of UU infection in the worse vagina sanitation was bigger.,compared with counterparts. UU is a potential risk factor in urinary-genital duct diseases. 4 UU biovar 1 dominated in the UU infection in our region. The UU biovar 2 infection in the population with urinary tract symptom and in females at the age of 16～35 were higher than those in no urinary tract symptom and in females at the age of 40～65 respectively. 5 UU to all of ten kinds of antimicrobial agents have resisted to some extent. The susceptibility rate of UU to Minomycin, Azithromycin, Doxycline, Joxamycin, Levofloxacin and Clarithromycin were higher , the rate to Erythromycin, Tetracycline, Ofloxacin were lower. Higher proportion of multidrug resistant existed. Chinese gall, chebulae, cortex phellodendri1 gardenia, paeoniae radix, et al exerted stronger inhibitory effect on the clinical UU strains.Traditional Chinese medicine, bazhengsan and wulinsan had the inhibitory effect on ureaplasma urealyticu to some extent. Complex prescription drug, liuheji,exerted stronger inhibitory effect on clinical UU isolates. 6 The correlation existed between tetM-positive and tetracycline phenotype- resistance in UU isolates, but not complete conformity. The rate of UU isolates carrying tetM was 39.02% totally, The rate of UU isolates carrying tetM was no difference between two UU biovars, but the rate in STD population was higher .