| Objectives: Urogenital tract infection caused by Chlamydia trachomatis(C.t)was one of the most common worldwide sexually transmitted diseases,the infection rate was more than Neisseria gonorrhoeae,and was in the first ranks of all sexually transmitted diseases,it appears gradually rising trend in China.Compared with the screening or antibiotic treatment,vaccination is more economical,and has a greater impact on global control of C.t infection.Therefore,vaccination is generally considered the best way to reduce C.t infection rates.At present,although there were lots of datas about C.t vaccine research,but so far there was no mature vaccine available.Currently focus of subunit vaccine and DNA vaccine research was to find the appropriate antigen,antigen studied comparatively was Major Outer Membrane Protein(MOMP)、Chlamydial Protease-Like Activity Factor(CPAF),however,these vaccines did not reach the desired immune effect.Polymorphic Membrane Proteins(Pmp)was recently discovered proteins located on the outer membrane of C.t,had advantages of larger molecular weight、specificity and conservative.Also,Pmp proteins may has important function on adhere to host cells and immune response after C.t infection.Therefore,Pmp protein was potential antigens of CT vaccines.So the purpose of this study was to investigate the polymorphism Of PmpF Gene Of Chlamydia Trachomatis causing urogenital tract infection,providing theoretical basis of Pmp F protein as Chlamydia trachomatis candidate vaccine.Methods: Clinics urogenital tract infection samples from three departments of urology,gynecology and dermatology in PLA 252 hospital from January 2013 to December 2014,patient’s related datas such as sex、age、source of departments was registered.First nucleic acid samples was screened with C.t nucleic acid diagnostic kit,positive samples was chosen to amplify PmpF using nested-PCR,then amplified PmpF gene was digested with four restriction endonucleases of Alu I,Hae III,Rsa I,Mob I,Restriction map was processed and analyzed Gelcompar II(version 3.5)image analysis software,UPGMA tree was obtained using Dice similarity coefficient and average linkage clustering method,the UPGMA tree can reflect polymorphism of PmpF gene.According to Pmp F-RFLP results,twenty six representative strains was selected to sequence and phylogenetic analysis,which can verify results of PmpF-RFLP.OmpI sequence of representative strains was amplifyed and sequence and compared to sequence of reference strain to determine serotypes of this strains.Relationship between genotype with patient gender,age and departments was explore using chi-square test.Results:1 From January 2013 to December 2014,178 C.t positive specimens was obtained from three departments of urology,gynecology and dermatology.In178 positive samples,105 specimens comes from men,accounting for 59%,73 comes from women,accounting for 41%;58 specimens comes from department of gynecology,accounting for 32.6%,88 specimens comes from department of urology,accounting for 49.4 %,32 specimens comes from department of dermatology,accounting for 18%;8specimens comes from patients less than 20 years of age,accounting for 4.5%,91 specimens comes from patients between 20 to 30 years old,accounting for 51.5%,59 specimens comes from patients between 30 to 40 years old,accounting for 33.1%,19 specimens comes from patients between 30 to 40 years old,accounting for 10.1%,only 1 specimens comes from patients between 30 to 40 years old,accounting for 0.6%.2 PmpF gene was amplified from 117 specimens in all 178 samples by Nested PCR,according to four kinds of restriction enzyme digestion pattern,117 PmpF gene can be divided into three Type,respectively as Type1,Type2,Type3.the number belong to Type 1 was 3,accounting for 2.6%,the number belong to Type2 was 43,accounting for 36.8%,the number belong to Type3 was 71,accounting for 60.7%,strains belong to Type3 was the most in the 3 types and was dominant strain in our study.26 representative strains was selected to sequence and phylogenetic analysis,results of phylogenetic analysis were consistent with the results of restriction enzyme digestion.3 The reslus of sequencing of OmpI of 26 representative strains showed that serotypes of these strains were D,Da,E,F,G,H,J respectively.4 Results of Chi-square test showed that age was correlated with type of PmpF gene,gender and department distribution was not correlated with type of PmpF gene.In patients less than 20 years of age,Type2 was the most,accounting for 75%;In patients betweent 20 to 30 years of age,Type3 was the most,accounting for 53.3%;In patients betweent 30 to 40 years of age,Type3 was aslo the most,accounting for up to 82.9%;In patients betweent 20 to 30 years of age,Type3 was the most,accounting for 57.1%.Conclusions:1 178 Urogenital C.t positive specimens was obtained from three departments of urology,gynecology and dermatology in our hospital,which shows that Chlamydia trachomatis has become common pathogens causing urogenital tract infections in this region.The age of infected patients was between 20 to 50 years,patients between 20 to 30 years old was the most.2 Serotypes of selected Sequenced strains were D,Da,E,F,G,H,J respectively,which was consistent with eported results.3 117 C.t strains can be divided into three type based the reslut s of restriction digestion of four restriction endonucleases of Alu I,Hae III,Rsa I,Mob I.strains belong to Type3 accounted for 60.7% and was dominant strain in our study.Results of phylogenetic analysis were consistent with the results of restriction enzyme digestion.4 Type of PmpF gene was correlated with age and not correlated with gender and department distribution.In patients less than 20 years of age,Type2 was the most;In patients more than 20 years of age,Type3 was the most... |
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