1256 2008 2011 - PCR-RFLP 4 2388 365 2.2 1 21~40 137 52 38.0% E 48 D 4 E Prevalence of chlamydia trachomatis urogenital tract infection and genotyping in out-patients of sexually transmitted disease clinics: a pilot study WANG Yan, FENG Wen-li, YANG Jing, WANG Yan-qing, JI Ying, YANG Hui-qing, ZHANG Rong-li. Department of Dermatology & Venerology, Second Hospital of Shanxi Medical U- niversity, Taiyuan 030001, China Abstract Objective To investigate Chlamydia trachomatis infection and genotypes in out-patients of sexually transmitted disease (STD) clinics in Taiyuan. Methods We collected the urethral or cervical secretion of out-patients with susceptible urogenital tract C. trachomatis infection, between 2008 and 2011, for examination of C. trachomatis by using ClearviewTM antigen assay kit (UK). A subgroup of positive specimens were subjected to culturing and genotyping via polymerase chain reaction-restriction fraction length polymorphism. Results Of the 2387 specimens within the four years, 365 tested positive to C. trachomatis antigen. There was a trend towards reduced annual incidence of infection among the population, with a male-to-female ratio of 2.2:1 and peak age of onset of 21-40 years. Of 137 clinical specimens sent for C. trachomatis culturing, 52 strains (48 of type E and 4 of type D) tested positive, thus resulting in a positive rate of 38.0%. Conclusion A comparatively high incidence rate of urogenital tract C. trachomatis infection, mostly genotyped as type E, in out-patients of STD clinic in Taiyuan deserves further large-scale study. Key words Chlamydia, trachomatis; Menbrane proteins; Genes; Polymorphism, restriction fragment length Chlamydia trachomatis Ct major outer membrane protein MOMP 19 A B/Ba C Ct L1 L2/L2a L3 D~K 2008 2011 non-gonorrhoea urethriti NGU human papilloma virus HPV Ct human immunodeficiency virus HIV - PCR- 200817 02200804 Email fwl92@hotmail.com 030001 RFLP Ct 1
1257 1.1 2008 2011 32 3000 r/min 60 min NGU 2 h 3 ml 60~ 1 72 h 1 ml 2SP -70 2 0.9% 2~3 90% -80 1.6 DNA 100 μl 1 ml Eppendorf 12 000 r/min 20 min 50 mmol/l KCl 2.5 mmol/l MgCl 2 10 mmol/l Tis-HCl ph8.3 0.1 g/l 0.45%NP-40 0.45% 20 200 mg/l CO 2 37 48 h K 55 1 h 3 15 min K 12 000 r/min 3 min -20 1.7 PCR-RFLP omp1 MOMP CO 2 35~37 24~48 h 1077 bp 5 -ATGAAAAAACTCTTGAAATCGG- 3 5 -TTTCTAGACTTCATCTTGTT-3 4 PCR 2 >4 5 6 Taq PCR Master Mix 12.5 μl DNA 5 μl 2 1.2 s 46 60 s 72 80 s 35 72 UNIPATH Clearview Ct 10 min 5 μl PCR 1% McCoy L Ct E 10 TBuffer BSA omp1 PCR Alu Msp MEM 37-20 1.3 8% 50 bp DNA Marker 2~3 cm 80 V 1.5 h 1~2 cm 30 s EB Ct D~ 1 K PCR-RFLP DNA 2SP Eppendorf 1.5 ml omp1-80 1.8 SPSS13.0 1.4 Ct Ct χ 2 P<0.05 >0.7 0.4~0.7 Ct 2.5 μl 94 5 min 94 50 <0.4 1.5 Ct McCoy 2 2.1 Ct 6-80 2008 2011 2388 Ct 2 6 2 1 87 38.4 ml DEAE-D 30 min 2 2.1.1 4 365 1 min 3000 r/min 15.3% Ct 5 min 5 ml χ 2 =35.319 P<0.01 1
1258 1 2008 2011 % 2008 692 150 21.7 2009 736 89 12.1 2010 584 88 15.1 2011 375 38 10.1 2387 365 15.3 0.119 Ct 0.146 Ct 20 40 18 22 Ct 18 7 11 1 1 22 16~ 2.1.2 2 1 19 1620 252 768 113 21~ 31~ 15.9% 15.6% 14.7% 19.5% 19.6% 12.5% 31~ 2.2 1 Ct 19.5% 21~ 19.6% 41~ 0.096 Ct 6.8% 8.9% 51 15.2% 21~ 41~ 51 14.3% Ct 2 / % % 0~20 18 1 5.6 22 0 0 21~ 541 86 15.9 326 64 19.6 31~ 657 128 19.5 273 34 12.5 41~ 292 20 6.8 112 10 8.9 51~ 112 17 15.2 35 5 14.3 1620 252 15.6 768 113 14.7 2.2 Ct 365 137 52 38.0% 137 40 44.4% 12 25.5% omp1 1077 bp 1 1 2 3 4 M E 5-100 bp -250 bp -500 bp -750 bp -1000 bp M D2000 1~5 E E 98% 3 M E 1 2 3 M 50bp DNA ladder E E 1~3 E 1 Ct omp1 Ct PCR-RFLP E 48 2 D 4 3 Ct BLAST omp1 CDC 2008 BLAST www.ncbi.nlm.nih.gov/blast Ct Ct 120 Ct E/bour DQ064286 D/B-185 X62919 280 6 2 Ct E
1259 3 M 1 2 M 50bp DNA ladder 1 2 D Ct D 9 2001 2003 1997 2000 10 2.2 1 Ct Ct 20 20 21~40 9 31~ 21~ ECDC 2007 Ct 25 122.6/10 41~ 51 5%~10% 2 Ct 2007 Ct PCR-RFLP Ct 2008 105 omp1 5.85% omp1 Ct 2.5 32.29/10 1.1 kb PCR 5 20.69% 3 Ct 100% 2008 2011 Ct 2388 Ct 365 15.3% 137 4.3%~20.3% 4-6 52 38.0% 76.9% 4.7% ~ 11 Ct 28.15% 4 7 D~K 2%~15% 12 13 Ct UNIPATH Mossman 14 Ct Ct EIA H 15 D/Da J/Ja 16 PCR E RFLP D 48% E F 17 PCR PCR PCR-RFLP E 27.9% F PCR 23.5% G 12.4% D 11.1% E 8 32.1% 27.1% F 18 28.3% Ct 2008 2011 E Ct 2 E D Ct Ct E
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