PURPOSE: The detection of high-level tetracycline-resistant strains of Neisseria gonorrhoeae (TRNG) can make important epidemiological contributions that are relevant to controlling infections from this pathogen. In this study, we aimed to determine the incidence of TRNG isolates over time and also to investigate the characteristics and genetic epidemiology of these TRNG isolates in Korea. MATERIALS AND METHODS: The antimicrobial susceptibilities of 601 isolates of N. gonorrhoeae from 2004 to 2011 were tested by standard Clinical and Laboratory Standards Institute methods. To determine the molecular epidemiological relatedness, N. gonorrhoeae multi-antigen sequence typing was performed. RESULTS: The incidence of TRNG increased from 2% in 2004 to 21% in 2011. The minimum inhibitory concentration distributions of ceftriaxone and susceptibility of ciprofloxacin in TRNG were different from non-TRNG and varied according to the year of isolation. Most of the TRNG isolates collected from 2004 to 2007 exhibited genetic relatedness, with sequence type (ST) 1798 being the most common. From 2008 to 2011, the STs of the isolates became more variable and introduction of genetically unrelated TRNG were noted. CONCLUSION: The increased incidence of TRNG strains until 2007 appears to be due, at least in part, to clonal spread. However, we propose that the emergence of various STs since 2008 could be associated with foreign import.
Anti-Bacterial Agents/*pharmacology ; Ceftriaxone/pharmacology ; Ciprofloxacin/pharmacology ; DNA, Bacterial/analysis ; Drug Resistance, Multiple, Bacterial/*genetics ; Gonorrhea/drug therapy/epidemiology/microbiology ; Humans ; Incidence ; Microbial Sensitivity Tests ; Molecular Epidemiology ; Neisseria gonorrhoeae/*drug effects/*genetics/isolation & purification ; Republic of Korea/epidemiology ; Sequence Analysis, DNA ; Tetracycline/pharmacology ; Tetracyclines/*pharmacology

OBJECTIVETo investigate the genotype and characteristics of Neisseria gonorrhoeae (N.gonorrhoeae) isolates with reduced susceptibility to ceftriaxone in Shenzhen from 2009 to 2011.

METHODSA total of 296 N.gonorrhoeae isolates were collected in Shenzhen from 2009 to 2011.ceftriaxone strains (minimum inhibitory concentration between 0.06 and 0.50 µg/ml) were determined by agar dilution method.Logistic regression was used to analyze the associated factors of ceftriaxone N.gonorrhoeae infection.Neighbor-joining (NJ) phylogenetic tree analysis and N.gonorrhoeae multi antigen sequence typing (NG-MAST) were performed on all ceftriaxone isolates and susceptible control isolates randomly selected in accordance with the principle of 1: 1 sampling.

RESULTSNo isolates displayed resistance to ceftriaxone, whereas 53(17.9%) showed reduced susceptibility to ceftriaxone among 296 isolates.Only antibiotic use in recent two months was associated with ceftriaxone isolates infection (OR = 3.080, 95%CI: 1.376-6.894) . Among the ceftriaxone isolates, 48 different ST were identified including 5 STs (ST1768, ST3927, ST641, ST7076 and ST7078) containing 2 isolates and 43 single STs. There were 26 STs previously reported from HongKong in China.Low sensitive strains clustering was not observed by NJ phylogenetic tree.

CONCLUSIONThe proportion of ceftriaxone strains among the 296 N.gonorrhoeae isolates collected from 2009 to 2011 in Shenzhen is high. The STs of ceftriaxone strains may have unique epidemic features in Shenzhen.

Adult ; Anti-Bacterial Agents ; pharmacology ; Ceftriaxone ; pharmacology ; China ; epidemiology ; Drug Resistance, Bacterial ; genetics ; Female ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; isolation & purification ; Young Adult

OBJECTIVETo identify the relationship between amino acid mutations in Neisseria gonorrhoeae isolates and their antibiotic resistance.

METHODSPI gene fragments of Neisseria gonorrhoeae from 17 clinical isolates were obtained with PCR amplification. They were cloned into the PCR cloning vector pBS-T to form pBS-T-PI and sequenced. The sequences of PI genes were analyzed. At the same time, minimum inhibitory concentration (MIC) of penicillin and tetracycline to these 17 isolates were measured and contrasted with the corresponding PI sequence.

RESULTSThe recombinants of PI gene from 17 clinical isolates of Neisseria gonorrhoeae were successfully constructed and sequenced. They were divided into PIA and PIB subtypes according to the results from blastn software by comparing the sequences with the GenBank. Mutations were found at the sites of 120 and 121. There were only some of the sequences having an aspartic acid (D) mutation on 120 and 121 sites, which was not the same as reported. On the other hand,there were two PI sequences,5-9 and 6-1, whose mutations on No. 120 were lysine, similar to those documented.

CONCLUSIONSome relationship between PI amino acids mutations at sites 120 and 121 in Neisseria gonorrhoeae isolates from Chengdu, China and their resistance to penicillin and tetracycline were found. However,further studies need to be done in the future to confirm this hypothesis.

Amino Acid Sequence ; Anti-Bacterial Agents ; pharmacology ; DNA Mutational Analysis ; DNA, Bacterial ; Drug Resistance, Bacterial ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Polymerase Chain Reaction

OBJECTIVETo investigate the blockness effects of purified polyclonal anti-porin I antibody on N. gonorrhoeae adherence to genitourinary tract epithelia of BALB/c mouse.

METHODSPolyclonal anti GST-PI antibody was generated by immunizing rabbit with GST-PI fusion protein which was constructed and expressed by ourselves. The purified immunoglobulin G was obtained by ammonium sulphate deposition and DEAE cellulose chromatography. Mice model of gonorrhea was established. In order to evaluate the effects of PI-IgG on gonococcus adhesion to vagina mucus, the macroscopic and pathological assessing as well as gonococcus culture was employed after gonococcus challenge on PI-IgG immunized mice.

RESULTNo pus and pathological inflammation were observed on mice vagina mucus treated with 1 mg/ml PI-IgG 3 hours before gonococcus challenge. Gonococcus could not be detected in the smears and washing solutions from vagina. Pathological inflammation was found in mice treated with anti PI-IgG, in which the concentrations were lower than 1 mg/ml or the treated time was longer than 3 hours prior to gonococcus challenge.

CONCLUSIONThe purified anti PI-IgG can effectively inhibit the adherence and infection of gonococci to genitourinary tract epithelia of BALB/c mice. In addition, the blocking duration of anti PI-IgG is associated with antibody concentration.

Animals ; Antibodies, Monoclonal ; pharmacology ; Bacterial Adhesion ; drug effects ; Epithelium ; drug effects ; microbiology ; Female ; Glutathione Transferase ; biosynthesis ; genetics ; Gonorrhea ; microbiology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Neisseria gonorrhoeae ; drug effects ; physiology ; Porins ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Urogenital System ; drug effects ; microbiology

OBJECTIVETo investigate the relationship between penA/ponA and penicillin resistance of Neisseria gonorrhoeae.

METHODSAgar dilution method was used to determine the minimum inhibitory concentrations (MICs) of the strains. Polymerase chain reaction-single stand conformation polymerphism (PCR-SSCP) and PCR- restriction fragment length polymorphism (RFLP) were used to detect the mutations in ponA and penA genes, which encoding the penicillin binding protein-1 and -2 (PBP1 and PBP2), respectively.

RESULTSAll the 80 N. gonorrhoeae isolates had a D345 insertion detected in penA while 93.7% of N. gonorrhoeae isolates having a point mutation Leu421 --> Pro in ponA. Most of the penicillinase producing N. gonorrhoeae (PPNG) strains possessed the mutations in ponA and penA.

CONCLUSIONOur data suggested that the plasmid and chromosome mediated penicillin-resistance conjugately increased the level of resistance.

Amino Acid Sequence ; Bacterial Proteins ; genetics ; Base Sequence ; DNA Mutational Analysis ; DNA, Bacterial ; Genes, Bacterial ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Mutagenesis, Insertional ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; Penicillin Resistance ; genetics ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo study the phenotypic and genotypic resistance to Fluoroquinolones in Neisseria gonorrhoeae (NG) isolated in Jiangsu province of China.

METHODSIn-vitro, susceptibility testing of ciprofloxacin and levofloxacin against ninety-five clinical isolates were determined by agar dilution method. Detection of mutation in the gyrA and parC genes was performed by polymerase chain reaction (PCR) assay and sequence analysis.

RESULTSThe clinical isolates demonstrated 100% resistance to ciprofloxacin. Based on gyrA and parC mutations, 18 types could be categorized among the 54 isolates. Based on the same gyrA mutations,isolates with high MIC appeared to have had more mutations in parC gene.

CONCLUSIONThe status of resistance to ciprofloxacin in NG was quite serious, and ciprofloxacin treatment for the treatment of NG infections in Jiangsu province should not be recommended. The results from this study suggested that mutations in the parC gene had contributed to the development of high Fluoroquinolone resistance in NG.

China ; DNA Gyrase ; genetics ; DNA Topoisomerase IV ; genetics ; Drug Resistance, Bacterial ; Fluoroquinolones ; pharmacology ; Genotype ; Gonorrhea ; drug therapy ; Humans ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Phenotype

OBJECTIVETo study the effect of compound nonoxinol on the inhibition of bacteria in vitro.

METHODSThe minimal bactericidal concentration (MBC) and minimal bactericidal time (MBT) of nonoxinol providine iodine and compound nonoxinol, composed of nonoxinol and providine iodine were determined by inhibiting C. albicans and N. gonorrhoeae.

RESULTSNonoxinol alone showed no inhibiting effects on bacteria. The MBT was 2 minutes for both compound nonoxinol and providine iodine, but the MBC of providine iodine was higher than that of compound nonoxinol.

CONCLUSIONCompound nonoxinol works better in inhibiting bacteria than either nonoxinoother or providine iodine used alone.

Anti-Infective Agents, Local ; pharmacology ; Candida albicans ; drug effects ; Dose-Response Relationship, Drug ; Drug Combinations ; Neisseria gonorrhoeae ; drug effects ; Nonoxynol ; pharmacology ; Povidone-Iodine ; pharmacology

OBJECTIVEUsing molecular epidemiology methods to investigate relationship between genotypes and drug-resistance of neisseria (N.) gonorrhoeae in Shanghai area.

METHODSA random amplified polymorphic DNA (RAPD) fingerprint method at the molecular level was used to differentiate the strains which were isolated from the outpatients of sexually transmitted disease clinics. The sensitivity to antibiotic of the 78 N. gonorrhoeae strains on 9 different antibiotics was tested and the relationship between different genotypes and phenotypes was studied.

RESULTSSelected RAPD primer could give out a group of amplification polymerase chain reaction bands with some main segments common to all the N. gonorrhoeae strains tested and some segments were different among the N. gonorrhoeae strains. All the 78 N. gonorrhoeae strains could be classified as three different groups (I, II and III). The strains could also be distinguished as four types (A, B, C and D) according to drug-resistance status. Using correspondence analysis method, the relationship between the three genotypes and four resistance types could be identified.

CONCLUSIONRAPD fingerprint seemed a useful genotyping method and could be used for molecular epidemiological studies.

Adolescent ; Adult ; Anti-Bacterial Agents ; pharmacology ; China ; epidemiology ; DNA Fingerprinting ; DNA, Bacterial ; genetics ; Drug Resistance, Bacterial ; Female ; Genotype ; Gonorrhea ; epidemiology ; microbiology ; Humans ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Molecular Epidemiology ; Neisseria gonorrhoeae ; classification ; drug effects ; genetics ; Random Amplified Polymorphic DNA Technique

OBJECTIVESTo investigate the correlation between mutation of GyrA and ParC genes and quinolone resistance in Neisseria Gonorrhoeae.

METHODSThe gene fragments of quinolone resistance-determining region (QRDR) in GyrA and ParC genes in 20 Neisseria gonorrhoeae strains clinically isolated in Wuxi, China, were sequenced, and the susceptibility of the 20 strains to quinolone was examined by agar diffusion method.

RESULTSThe mutations at the Asp95 point in GyrA gene were found in 20 strains. Of the 19 stains examined, 16 had mutations at the 86, 87, 88, 91 points in ParC genes.

CONCLUSIONSThe mutations of Asp95 in GyrA gene and Asp86, Ser87, Ser88, Glu91 in ParC gene contribute to quinolone resistance in Neisseria Gonorrhoeae.

Base Sequence ; DNA Gyrase ; genetics ; DNA Topoisomerase IV ; genetics ; Drug Resistance, Bacterial ; genetics ; Humans ; Molecular Sequence Data ; Mutation ; Neisseria gonorrhoeae ; drug effects ; genetics ; isolation & purification ; Quinolones ; pharmacology

OBJECTIVETo investigate the antimicrobial susceptibility,auxotype, and plasmid profile of Neisseria gonorrhoeae isolates in China and to provide evidence for the development of treatment guideline and policy for control.

METHODSAgar dilution was used to detect antimicrobial susceptibility. The auxotype was determined by GC genetic medium. The plasmid was extracted by alkaline cleavage and electrophoresed.

RESULTSA total of 4,976 gonococcal isolates were tested in the last 8 years. The resistant rate for penicillin was 71.60% with PPNG being 15.54%. Tetracycline-resistant (TRNG) isolates accounted for 93.02% with 10.48% high level tetracycline-resistant. The resistant rate for ciprofloxacin was also relatively high (31.78%). The resistant rates for spectinomycin and ceftriaxone were 0.36% and 0.46%. The predominant auxotypes of gonococcal isolates were proto and pro(-) during 1995 - 1996 in Nanjing, accounted for 46.4% and 47.53%, 48.4% and 50.22%, respectively. There were 8 strains harboring 4.2, 5.4, 39.5 kb plasmids and 2 harboring 4.2, 4.9, 5.4, 39.5 kb plasmids in 10 PPNG strains; 2 harboring no plasmid, 28 harboring 4.2, 4.9, 5.4, 39.5 kb plasmids in 30 non-PPNG strains. The 5.4 kb plasmid of PPNG could be digested with restriction endonuclease BamHI while the 5.4 kb plasmid of non-PPNG could not.

CONCLUSIONThe gonococcal isolates were highly resistant to penicillin, tetracycline, and ciprofloxacin, while were still sensitive to spectinomycin and ceftriaxone. No significant auxotyping change was found in terms of predominant gonococcal strains in the last two years in Nanjing while 5.4 kb plasmid might be the most prevalent resistant plasmid in Nanjing.

China ; Drug Resistance, Bacterial ; Microbial Sensitivity Tests ; Neisseria gonorrhoeae ; drug effects ; isolation & purification ; Time Factors