|
Chlamydia trachomatis (Chlamydia trachomatis, Ct) infection is the most common sexually transmitted pathogens, can cause urethritis, cervicitis, endometritis, epididymitis, prostatitis, infertility variety of diseases, and the incidence trend of yearly The upward trend has become a great harm to public health problems. Currently, the incidence of urogenital Ct infection and its prevalence is closely related to antibiotic therapy is an important disease secondary prevention strategy; Ct vaccine still can not be used in clinical antibacterial drugs for the treatment of Ct infection becomes more important. However, the clinical manifestations of patients with urogenital Ct infection is often non-specific or in the form of occult, not easy to pay attention, resulting in a protracted illness, relapse or recurrent infections, Ct reduced sensitivity to a variety of antibiotics, clinical treatment failure cases dramatically increase. Objective: To detect the urogenital tract standard strains and clinical isolates of C. trachomatis alone azithromycin, minocycline, moxifloxacin, doxycycline and rifampin five clinical antibiotics commonly used in vitro and joint drug sensitivity, research and drug clinical efficacy and tolerability of molecular biology mechanism. : Collection 2005-2009 to Tianjin Sexually Transmitted Disease Institute, outpatient and patients meet the sampling criteria between male urethra or female cervical epithelial cells, and records related to the clinical information. McCoy cell culture method cultured clinical strains, all clinical strains of five generations of blind passage culture positive samples continue subculture until the infection rate is more than 90%, a total of 41 clinical isolates. Standard strains and 41 clinical strains were used to separate and combined sensitivity detection microdilution method and checkerboard dilution method of five commonly used antibacterial drugs. Nucleic acid amplification omp1 gene of clinical strains and genotyping Alu Ⅰ, Msp Ⅰ double digestion. Clinical treatment failure cases of clinical strains, the PCR method for detection of macrolide resistance-associated 23S ribosomal RNA gene products were sequenced to detect mutations; restriction fragment analysis (RFLP) detection gyrA quinolone resistance determining region (Quinolone-Resistance Determining Region, QRDR) common point mutation. Statistical Methods: one-way ANOVA with SPSS11.5 software (One-Way ANOVA), significant in the analysis of variance, require data to meet the homogeneity of variance pairwise comparisons using the most significant difference method that LSD (least-significant different) method for multiple comparisons between groups. P lt; 0.05 for the difference was statistically significant. Results: A total of 238 cases develop clinical specimens, 41 patients with positive clinical strains, the positive rate of about 17.23%. Susceptibility alone, this 41 specimens for detection of the minimum inhibitory concentration (MIC) (Unit ug/m1) are: Azithromycin 0.063-0.5 moxifloxacin 0.03-0.24, the minocycline 0.008-0.064, doxycycline prime 0.063-0.5, rifampicin 0.002-0.016. United susceptibility testing inhibitory concentration (FIC) shows: azithromycin and moxifloxacin, doxycycline, rifampin joint in vitro, respectively, 51.22%, 53.66% and 58.54% of the strains of a synergistic or additive effect less antagonism; ANOVA test indicated no statistically significant difference between the three groups (when a = 0.05, P = 0.755). Joint in vitro, minocycline and azithromycin, moxifloxacin, rifampin, respectively, 90.24%, 85.37%, 92.68% Ct strains antagonism synergies. Clinical treatment failure the strains corresponding resistance gene was not detected. Conclusion: 1, successfully isolated and cultured Ct clinical strains, to lay the foundation for further development of the pathogenic mechanism of the pathogens, immune mechanisms, in vitro drug susceptibility testing, resistance mechanisms and protective vaccine research. 2, Ct showed varying degrees of decreased sensitivity to commonly used antimicrobial drugs, MIC value is higher than those of the past, reported in the literature. Research on the relationship between Ct resistance in vitro antibacterial effect is not clear. 3 different joint susceptibility testing of antimicrobial agents in vitro, to some extent, be able to make up for some shortcomings of individual susceptibility test with significant clinical significance and will further study the combined effects of the mechanism of antimicrobial drugs experimental basis.
|