RESUMO
BACKGROUND: In the present study, we characterized multidrug-resistant Pseudomonas aeruginosa (MDRP) clinical isolates from a paediatric facility and investigated the types and features of the metallo-beta-lactamases (MBLs) produced by carbapenem-resistant strains. METHODS: Four hundred and ninety-eight strains of Pseudomonas aeruginosa were isolated from patients at Beijing Children's Hospital between January 2005 and December 2006. The minimal inhibition concentrations (MICs) of the strains for 13 antibiotics were measured. A combination of the E test and PCR amplification/DNA sequencing was used to define the carbapenem-resistant strains. RESULTS: We found that 24.1% (120/498) of the isolates were MDRP. The frequencies of resistance to imipenem and meropenem were 34.2% and 35.8%, respectively, and the MIC50 and MIC90 values for the two antibiotics were identical at 4 microg/ml and 32 microg/ml, respectively. The detection rate for carbapenem resistance was 49.2% (59/120). Among the 59 carbapenem-resistant Pseudomonas aeruginosa strains, 39 (66.1%) were positive for the MBL genotype; 35 (89.7%) strains carried the bla(IMP) gene and 4 (10.3%) strains carried the bla(VIM) gene. Neither bla(SPM) nor bla(GIM) was amplified from any of the 59 isolates. DNA sequencing revealed that IMP-1 was present in 35 IMP-producing isolates and VIM-2 was detected in four VIM-producing isolates. CONCLUSIONS: These MDRP isolates exhibited high frequencies of resistance to carbapenems among clinical isolates from a paediatric facility in Beijing, China. The production of MBL appears to be an important mechanism for carbapenem resistance in Pseudomonas aeruginosa.
Assuntos
Farmacorresistência Bacteriana Múltipla , Pseudomonas aeruginosa/efeitos dos fármacos , beta-Lactamases/biossíntese , Carbapenêmicos/farmacologia , Criança , Humanos , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa/enzimologia , Análise de Sequência de DNA , beta-Lactamases/classificação , beta-Lactamases/genéticaRESUMO
OBJECTIVE: To analyze the antibiotic resistance of the Pseudomonas aeruginosa (PA) isolated from pediatric patients and the resistant genes of beta-lactam antibiotics thereof. METHODS: 146 PA strains were isolated from pediatric patients. Agar dilution method recommended by the Clinical and Laboratory Standards Institute was used to examine the minimum inhibitory concentrations (MICs) of 12 antimicrobial agents, including the penicillins, third and fourth genet ration cephalosporins, carbapenemase, Aztreonam, beta-lactamase inhibitors, quinolones, and aminoglycosides. PCR was used to detect the expression of the genes TEM, SHV, OXA, PER, GES, CTX-M, IMP, VIM, DHA, MIR, FOX, and oprD2. RESULTS: The multi-drug resistance rates against different antibiotic were high among the 146 PA strains. The rates of imipenem and meropenem resistance were 41.1% and 35.6% respectively. Among the 146 PA strains, 46 (31.5%) were positive for the MBL genotype; 38 (82.6%) carried the blaIMP gene, 8 (17.4%) carried the blaVIM gene, and 114 (78.1%) were oprD2 negative. The genes TEM, SHV, OXA, CTX-M, PER, VEB, GES, FOX, MIR, and DHA were not found in all strains. CONCLUSION: Many PA isolated from pediatric patients carry the genes IMP or VIM and losses oprD2 gene related to the expression of the outer membrane porin OprD2. The loss of the gene oprD2 is essential mechanism of beta-lactam antibiotics resistance in PA.
Assuntos
Genes Bacterianos , Pseudomonas aeruginosa/genética , Resistência beta-Lactâmica/genética , Proteínas da Membrana Bacteriana Externa/genética , Criança , Farmacorresistência Bacteriana Múltipla/genética , Humanos , Testes de Sensibilidade Microbiana , Porinas/genética , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/isolamento & purificaçãoRESUMO
OBJECTIVE: To study the drug resistance profiles of extended spectrum beta lactamase (ESBL) producing Escherichia coli (E. coli) and provide guidance for pediatric clinical prescription. METHODS: 5127 strains of ESBL producing E. coli were isolated from 5 children's hospitals in Beijing, Shanghai, Chongqing, and Guangzhou. The minimum inhibitory concentration (MIC) values of 349 strains to 9 antibiotics most in use in pediatrics were measured with agar dilution method. RESULTS: The prevalent rate of ESBL producing of E. coli was 46.7%. The resistant rates to ampicillin/clavulanic acid, cefotaxime, ceftazidime, and cefepime were 23.1%, 67.2%, 24.5%, and 48.4% respectively, and the intermediate rates were 38.5%, 26.4%, 5.7%, and 19.1% respectively. The resistant rate to amikacin was 5.4%. All the strains were susceptible to imipenem. The MIC90 values of ampicillin, cefotaxime, cefepime, ciprofloxacin, and gentamicin were 256 mg/L or higher. There were differences in the resistance profile to ampicillin/clavulanic acid, cefepime, ciprofloxacin, and gentamicin among different regions. CONCLUSION: ESBL is extensively prevalent among the E. coli strains isolated from the pediatric clinic that are resistant to most antimicrobial agents except imipenem and amikacin. Regional difference exists in drug resistance to some agents exist.
Assuntos
Infecções por Escherichia coli/microbiologia , Proteínas de Escherichia coli/metabolismo , Escherichia coli/enzimologia , beta-Lactamases/metabolismo , Anti-Infecciosos/farmacologia , Criança , Pré-Escolar , China/epidemiologia , Farmacorresistência Bacteriana Múltipla , Escherichia coli/efeitos dos fármacos , Escherichia coli/metabolismo , Infecções por Escherichia coli/epidemiologia , Hospitais Pediátricos/estatística & dados numéricos , Humanos , Lactente , Recém-Nascido , Testes de Sensibilidade Microbiana , Prevalência , Resistência beta-Lactâmica , beta-Lactamas/farmacologiaRESUMO
OBJECTIVE: To investigate the antimicrobial resistance of Streptococcus pneumoniae (S.pneumoniae) isolated from Chinese children with pneumonia. METHODS: Hypopharyngeal aspirate specimens were collected from hospitalized children with pneumonia who were admitted to the children's hospital located in Beijing, Shanghai, Guangzhou or Shanghai from February 16, 2006 to February 16, 2007. The minimum inhibitory concentration (MIC) of S.pneumoniae isolates against penicillin, amoxicillin, cefuroxime (sodium), ceftriaxone, erythromycin, vancomycin, ofloxacin and imipenem was determined by E-test method. RESULTS: A total of 279 S.pneumoniae isolates were obtained. Eighty-six percent of the isolates were not susceptive to penicillin, and 23.3% was resistant to penicillin. The rate of susceptibility of the isolates to amoxicillin was 92.1%, and to cefuroxime and ceftriaxone was 19.0% and 75.3%, respectively. The isolates also showed a high susceptibility to vancomycin (99.6%) and ofloxacin (97.8%). Seventeen point six percent of the isolates were not susceptive to imipenem, and most of those were intermediate. Almost of all isolates were resistant to erythromycin. There were some distinct regional differences in the susceptibility to antimicrobials tested except for erythromycin, vancomycin and ofloxacin. CONCLUSIONS: The S.pneumoniae isolates from Chinese children with pneumonia were susceptive to amoxicillin, vancomycin and ofloxacin, but were not susceptive or resistant to penicillin, cefuroxime and erythromycin. The isolates kept susceptibility to ceftriaxone and imipenem to a certain extent.
Assuntos
Streptococcus pneumoniae/efeitos dos fármacos , Criança , Farmacorresistência Bacteriana , Hospitalização , Humanos , Testes de Sensibilidade MicrobianaRESUMO
OBJECTIVE: To study if the resistance to macrolide in Enterococcus can be transferred between strains, and species of the same and different genera. METHODS: Agar dilution was used to screen 30 enterococcal isolates that were resistant to erythromycin. Conjugation was performed by filter mating method. The 30 donor bacteria included 13 strains of Enterococcus faecalis, 16 strains of E. faecium, and 1 strain of E. hirae. The recipient bacteria included 1 strain of E. faecalis sensitive to erythromycin and resistant to tetracycline, and 1 strain of Staphylococcus aureus with the MIC against erythromycin of 0.25 approximately 1 microg/ml. Polymerase chain reaction was used to test the existence of ermB gene and the tranposons Tn1545 and Tn917 in the enterococcal isolates before and after filter mating. RESULTS: The transfer rate between different strains and species of the same genus were all 100%. The MIC(50) and MIC(90) against erythromycin of 13 conjugates were both 512 microg/ml, and Tn1545 and Tn917 were found in the ermB gene of 12 conjugates. 17 conjugates were obtained from 16 strains of donor E. faecium and 1 strain of E. hirae with the MIC(50) and MIC(90) both of 512 microg/ml. The ermB gene was found in 16 of the 17 conjugates, and 11 of the 16 conjugates showed the existence of Tn1545 and Tn917, Tn1545 existed in the ermB gene of 4 conjugates, and Tn917 existed in the ermB gene of 1 conjugate. 30 conjugates of Staphylococcus aureus were obtained by plasmid conjugation and transfer with a transfer rate of 100% and the MIC(50) and MIC(90) both of 512 microg/ml. The ermB gene was found in 28 of the 30 conjugates. Both Tn1545 and Tn917 were found in the ermB gene of 23 of the 28 conjugates, Tn1545 was found in the ermB gene of 4 conjugates, and Tn917 was found in the ermB gene of 1 conjugate. CONCLUSION: The resistance to macrolide of Enterococcus, related with the existence of ermB gene and transposons Tn1545 and Tn917, can be transferred between strains and species of same and different genera.
Assuntos
DNA Bacteriano/genética , Farmacorresistência Bacteriana/genética , Enterococcus/genética , Transferência Genética Horizontal , Antibacterianos/farmacologia , Conjugação Genética/genética , Enterococcus/efeitos dos fármacos , Enterococcus/isolamento & purificação , Eritromicina/farmacologia , Testes de Sensibilidade Microbiana , Plasmídeos/genética , Especificidade da EspécieRESUMO
OBJECTIVE: To investigate the carrier rate of Streptococcus pneumoniae and the susceptibility of S. pneumoniae to 10 different antimicrobial drugs among children in China. METHODS: Nasopharageal swab specimens were collected from randomly selected 3578 out-patients with upper respiratory infection aged 1 month to 5 years in Beijing, Shanghai, and Guangzhou 2000-2002 so as to isolate S. pneumoniae. The susceptibility of these strains to erythromycin and beta-lactam antibiotics (penicillin, amoxicillin/clavulanic acid, cefaclor, cefuroxime and ceftriaxone) was determinated by E-test MIC method, and the susceptibility to chloramphenicol, tetracycline, sulfamethoxazole/trimethoprim and ciprofloxacin was determinated by disk diffusion method. RESULTS: A total of 892 strains of S. pneumoniae were isolate from the 3578 children with a carrier rate of 24.9% (9.0%-36.0%). 634 isolates of S. pneumoniae were tested to examine their susceptibility to chloramphenicol, 199 to ciprofloxacin, and 887 to the other 8 antimicrobial drugs. The insusceptibility rate of S. pneumoniae to penicillin was 39.9%, and the resistance rate was 6.4%. The resistance rate to cefaclor was 20.4%. The insusceptibility rate to cefuroxime was 19.0%. The susceptibility rate to amoxicillin/clavulanic acid was 98.5% and that to ceftriaxone was 97.2%. More than 80% of the isolates were resistant to erythromycin, tetracycline, and sulphamethoxazole/trimethoprim. Chloramphenicol resistance was less common (32.3%) than the resistance to other non-beta-lactam antibiotics, except for ciprofloxacin to which the resistance rate was 2.0%. The national insusceptibility rates for beta-lactam antibiotics did not vary significantly during the three years with the widest varying range of 5.1% for penicillin (38.1%-43.2%). Each of the three cities had their different varying patterns of antimicrobial susceptibility, especially for beta-lactam antibiotics. Multi-drug resistance was common (88.7%). Strains of S. pneumoniae insusceptible to penicillin were significantly more common than those susceptible to penicillin (99.1% vs. 78.8%, chi(2) = 50.36, P < 0.001), and strains of S. pneumoniae insusceptible to erythromycin were significantly more common than those susceptible to erythromycin too (95.8% vs. 17.1%, chi(2) = 360.26, P < 0.001). CONCLUSION: Antimicrobial resistant S. pneumoniae has already become a serious problem in China. Ongoing surveillance study on the antimicrobial resistance of S. pneumoniae is necessary for appropriate antimicrobial use in clinical work and modification of medical strategies for people's health.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Infecções Respiratórias/microbiologia , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/isolamento & purificação , Pré-Escolar , Feminino , Humanos , Lactente , Masculino , Testes de Sensibilidade Microbiana , Infecções Pneumocócicas/microbiologiaRESUMO
OBJECTIVE: To investigate the situation of antibiotic resistance of Haemophilus influenzae isolates from Chinese children. METHODS: Eight hundred and ninety-eight strains of H. influenzae isolated from randomly selected pediatric out-patients in Beijing, Shanghai and Guangzhou 2000 approximately 2002 underwent determination of antibiotic susceptibility by E test MIC method for beta-lactam antibiotics (ampicillin, amoxicillin/clavulanic acid, ceftriaxone, cefuroxime, and cefaclor) and KB disc diffusion method for chloramphenicol, tetracycline, sulfamethoxazole/trimethoprim (SMZ/TMP), azithromycin, and ciprofloxacin. RESULTS: The average rate of beta-lactamase production among these isolates was 12%. The MIC50 and MIC90 of ampicillin were 0.25 microg/ml and 4 microg/ml respectively for the 898 isolates, 88% of the isolates were susceptible to ampicillin and 10.6% was resistant. There was no beta-lactamase negative strains among the ampicillin intermediately-susceptible and resistant strains. All the isolates were susceptible to amoxicillin/clavulanic acid, ceftriaxone, cefuroxime and azithromycin. 96.1% of the H. influenzae isolates generally remained susceptible to cefaclor with a MIC50 of 1.5 microg/ml and a MIC90 of 3 microg/ml, and 99% of the isolates were susceptible to ciprofloxacin. The susceptibility rate to tetracycline of the isolates from Beijing and Shanghai areas were 57% and 61% respectively, both lower than the susceptibility rates of the isolates from Guangzhou (81%), while the resistant rate to SMZ/TMP of the isolates from Beijing was 35%, lower than those of the isolates from Guangzhou (47%) and Shanghai (54%). The resistant rate to ampicillin was 7.7% in 2000 and was increased to 14.5% in 2002. 34.5% of all the isolates were susceptible to 8 antimicrobial agents while 12.8% was multi-drug resistant. 58.5% of the isolates were distributed in five major antimicrobial resistance profiles in which SMZ/TMP resistance (29.8%) was the most prevalent, followed by SMZ/TMP and tetracycline combined resistance (12.5%) and tetracycline resistance (9.4). The ampicillin non-susceptible isolates had higher resistant rates to cefaclor (23.5%), tetracycline (63.3%), SMZ/TMP (74.5%) and chloramphenicol (63.3%) than the ampicillin susceptible strains (1.6%, 51.4%, 11.1% and 4.9% respectively). CONCLUSION: Beta-lactamase production and ampicillin resistance among the isolates from Chinese children are increasing generally during the period 2000 approximately 2002. There is variation of resistance among different regions. There are strong correlations between ampicillin resistance and resistance to cefaclor, chloramphenicol, and tetracycline among the H. Influenzae isolates.
Assuntos
Ampicilina/farmacologia , Antibacterianos/farmacologia , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/efeitos dos fármacos , Resistência a Ampicilina , Pré-Escolar , Cloranfenicol/farmacologia , Ácido Clavulânico/farmacologia , Farmacorresistência Bacteriana , Feminino , Haemophilus influenzae/isolamento & purificação , Humanos , Lactente , Masculino , Testes de Sensibilidade MicrobianaRESUMO
OBJECTIVE: To investigate the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates from Chinese children in seven cities. METHOD: A total of 134 MRSA isolates were collected from nine hospitals. Multilocus sequence typing and spa typing were analyzed by polymerase chain reaction (PCR), and staphylococcal chromosomal cassette mec (SCCmec) type was analyzed by multiplex PCR. The Panton-Valentine leukocidin (pvl) gene was also detected. RESULT: Most MRSA strains were isolated from pneumonia and skin and soft tissue infection (SSTIs) patients, accounting for 82.1%. Overall, 16 sequence types (STs) were obtained, and CC59 (51.7%) was found to be the most prevalent, which included ST 59 and ST 338, followed by ST239 (16.4%). SCCmec types II, III, IV, and V were also identified in the current study. SCCmec type IV was the most predominant type at 50.0%, followed by SCCmec type V at 23.9% and III at 23.9%. SCCmec subtypes IVa, IVc, and IVg were found among SCCmec type IV strains, whereas IVa was the main subtype at 77.6%. Twenty-six spa types were also identified, among which the predominant type was t437 (47.8%). The prevalence of pvl genes and the SCCmec type of strain was relevant, and the pvl gene positive rate was higher in SCCmec type IV and V-type strains than in SCCmec type II and III strains (58.6% vs. 14.3%, P < 0.05); there was a significant difference between them. In the strains isolated from pneumonia and SSTIs, ST59-MRSA-IVa(t437) was the predominant clone. There were five clones detected from the strains isolated from septicemia, with ST59-MRSA-IVa(t437) and ST59-MRSA-V(t437) as the main clones (57.1%). Various predominant clones existed in different regions. ST59-MRSA-IVa(t437) was the prevalent clone in the Guangzhou, Beijing, Chongqing, and Shenzhen areas, whereas ST239-MRSA-III(t037) was the prevalent clone in the Shanghai area. Fifty percent of the isolates from the Wenzhou area belonged to ST910-MRSA-V(t318), whereas three clinical strains isolated from the Shenyang region belonged to three different types. CONCLUSION: The results indicate that MRSA isolates from Chinese children are largely associated with the ST59-MRSA-IV(t437) and ST239-MRSA-III(t037) clones. These two may belong to community-acquired MRSA and hospital-acquired ones, respectively. Different prevalent clones were detected in different diseases and different regions. Therefore, there is a need to conduct further research on clinical isolates, which can guide the choice of antibiotic treatment and the examination of MRSA prevalence.
Assuntos
Staphylococcus aureus Resistente à Meticilina/genética , Infecções Estafilocócicas/microbiologia , Adolescente , Técnicas de Tipagem Bacteriana , Criança , Pré-Escolar , China/epidemiologia , DNA Bacteriano/genética , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Staphylococcus aureus Resistente à Meticilina/classificação , Staphylococcus aureus Resistente à Meticilina/isolamento & purificação , Prevalência , Infecções Estafilocócicas/epidemiologiaRESUMO
BACKGROUND: Despite the prevalence of Streptococcus pneumoniae serotype 19A, the molecular characteristics of this serotype are yet to be fully elucidated. The aim of this study was therefore to determine the homology of the serotype 19A in China. METHODS: Pulsed-field gel electrophoresis and multilocus sequence typing were done to these forty-nine serotype 19A isolates to investigate the relationship between the strains prevalent in Beijing and other regions. RESULTS: From 1997 to 2006, the percentage of serotype 19A isolates increased. The susceptibility rate to penicillin and amoxicillin decreased and the resistance rate to cefuroxime increased. ST320 was the most prevalent ST, followed by ST3546. There were six new STs identified in our study. The serotype 19A strains were classified into six different pulsed-field gel electrophoresis (PFGE) patterns. ST320, which was associated with two different PFGE patterns (A and D), accounted for 32 isolates, and ST3546, which was associated with two PFGE patterns (B and E), accounted for eight isolates. CONCLUSIONS: From 2003 onwards, ST320 was the most common ST and the rate of resistance to cefuroxime increased significantly. Further long-term surveys of Streptococcus pneumoniae serotype 19A are required to monitor ST prevalence and antimicrobial resistance in this important human pathogen.
Assuntos
Infecções Pneumocócicas/microbiologia , Streptococcus pneumoniae/genética , Pré-Escolar , China , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Humanos , Lactente , Recém-Nascido , Epidemiologia Molecular , Infecções Pneumocócicas/tratamento farmacológico , Sorotipagem , Streptococcus pneumoniae/classificação , Streptococcus pneumoniae/efeitos dos fármacos , Streptococcus pneumoniae/isolamento & purificação , Fatores de TempoRESUMO
OBJECTIVE: The present study was designed to investigate the situation of serotype distribution and beta-lactam antibiotics resistance of Streptococcus pneumoniae (S. pneumoniae) isolated from Chinese children, and to further understand the significance of vaccine for preventing infection caused by the bactria and controlling the resistance to antibiotics. METHODS: Nasopharageal swab specimens were collected from randomly selected less than 5-year-old out-patients with upper respiratory infection in Beijing, Shanghai and Guangzhou, 2000 - 2002. Capsular typing was performed by the Quellung reaction tested using a simplified chessboard system for typing of S. pneumoniae. The coverage rate of the 7-valent pneumococcal conjugate vaccine (4, 6B, 9V, 14, 18C, 19F and 23F) was calculated. Antibiotic susceptibility was determined by E-test MIC method for beta-lactam antibiotics (penicillin, amoxicillin/clavulanic acid, cefaclor, cefuroxime and ceftriaxone). RESULTS: Totally 625 pneumococcal strains were typed. Serogroup 19, including 121 strains, was the most frequent serogroup observed (19.4%). Other frequently observed serotypes/serogroups in decreasing order of frequency were serotype/serogroups 23 (15.4%), 6 (13.3%), 14 (6.6%) and 15 (4.3%). Of all these isolates, about 57.6% (360/625) were in the 7-valent conjugate vaccine. Only 1, 6 and 12 strains were serotypes/serogroups 4, 9 and 18, respectively. The coverage rate for the 7-valent vaccine of penicillin nonsusceptible S. pneumoniae (PNSP) was higher than penicillin susceptible S. pneumoniae (PSSP) (73.2% and 46.1%). Serogroups 19 and 23, without other serotypes/serogroups, were significantly associated with PNSP (serogroup 19 accounted for 29.1% of PNSP and 12.2% of PSSP; serogroup 23 accounted for 23.8% of PNSP to 9.2% of PSSP). Overall, 140 strains (22.4%) could not be typed by using the chessboard system, and 117 strains (18.7%) were identified as other 28 kinds of serotype/serogroup. The strains showed different resistance change for beta-lactam antibiotics according to different serotype/serogroup during the three years. CONCLUSIONS: Serotype/Serogroup 19, 23, 6, 14 and 15 were the common types among the pneumococcal strains isolated from Chinese children. Serogroups 19 and 23 were significantly associated with PNSP. The 7-valent pneumococcal conjugate vaccine could cover most of the islotes.
Assuntos
Farmacorresistência Bacteriana Múltipla , Infecções Pneumocócicas/epidemiologia , Streptococcus pneumoniae/classificação , Pré-Escolar , China/epidemiologia , Humanos , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/microbiologia , Sorotipagem , Streptococcus pneumoniae/efeitos dos fármacosRESUMO
OBJECTIVE: To investigate the molecular epidemiology of the penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) in Beijing, China. METHODS: The resistant profile of 63 PNSP strains isolated from children with upper respiratory infection in the outpatient department from 2000 to 2002 was analyzed. The isolates were compared by detecting restriction fragment length polymorphism (RFLP) of the penicillin-binding protein (PBP) genes pbp1a, pbp2b and pbp2x and by applying chromosomal macrorestriction patterns detected by pulsed-field gel electrophoresis (PFGE). RESULTS: Sixty-one (96.8%) out of the 63 PNSP strains were multidrug-resistant Streptococcus pneumoniae (Sp). Overall, 16 resistance profiles were found, 14 of which were multidrug resistant profiles. Seven (33.3%), 6 (24.0%) and 8 (47.1%) strains resistant to one of cephalosporins were respectively isolated in 2000, 2001 and 2002, indicating an increasing trend but without any statistical significance (chi(2) = 2.42, P > 0.05). The RFLP results showed 8, 9 and 18 genotypes of pbp1a, pbp2b and pbp2x, respectively. A total of 30 patterns were found according to the three pbps types. And clearly, the most common 5 patterns had main resistant profiles. In the mean time, 35 different PFGE types were elucidated and the 9 PFGE types, with each consisting of more than 2 strains, covered 59% (37/63) of all isolates. One of the 9 PFGE type included 2 strains, both possibly related to each other, but one of them was detected to be the same PFGE pattern with clones prevalent in Asia, Vietnam-19 serogroup, Singapore-19 serogroup, Taiwan-19 serogroup, and the other was the same as that in Korea-19 serogroup. CONCLUSION: Multidrug resistance is very common among PNSP isolates in Beijing. The spread of a few multidrug resistant clones is an important factor for the prevalence of PNSP. It deserves the concern that the resistant clones spread in Asia have been found in Beijing.
Assuntos
Farmacorresistência Bacteriana/genética , Resistência às Penicilinas/genética , Infecções Pneumocócicas/epidemiologia , Streptococcus pneumoniae/genética , Pré-Escolar , China/epidemiologia , DNA Bacteriano/genética , Genótipo , Humanos , Lactente , Epidemiologia Molecular , Polimorfismo de Fragmento de Restrição , Análise de Sequência de DNA , Streptococcus pneumoniae/classificaçãoRESUMO
OBJECTIVE: Haemophilus (H.) influenzae is a gram-negative bacillus that is a common commensal organism of the human upper respiratory tract and an important cause of human diseases such as pneumonia, meningitis, septicemia, epiglottitis and cellulitis. Strains of H. influenzae are classified according to their capsular polysaccharide. There are six serotypes, designated as a through f. In addition, there are nonencapsulated strains. Although the type of infectious diseases caused by H. influenzae has changed considerably in recent years because of the widespread and routine immunization of children against type b H. influenzae (Hib), Hib remains an important pathogen. Ampicillin is the drug of choice for treating many infections caused by H. influenzae, but its usefulness has been compromised by the increasing prevalence of ampicillin-resistant strains. The continued monitoring of resistant strains by using genotyping methods may provide insights into the epidemiology of transmission. A molecular epidemiological study of ampicillin-resistant H. influenzae derived from nasopharyngeal swabs specimens of children less than 5 years of age with respiratory tract infection were investigated in this study. METHODS: A total of 899 isolates were collected from Beijing, Shanghai, and Guangzhou during 2000-2003. Susceptibility to ampicillin was determined by using E-test. Ampicillin-resistant H. influenzae strains were selected according to National Committee for Clinical Laboratory Standards (NCCLS) 2002 breakpoints. Nested PCR method with primers specific for bexA gene and b capsulate type-specific gene was established. Genotyping by pulsed-field gel electrophoresis (PFGE) and multiplex PCR assay was performed for all ampicillin-resistant H. influenzae strains. RESULTS: Seventy-four ampicillin-resistant H. influenzae strains were obtained. Two strains were positive by nested PCR, characterized as b genotype. The incidence of Hib in ampicillin-resistant H. influenzae strains was 2.7%; 38 genotypes were detected by PFGE. Detection of five types strains of clonal dissemination by PFGE accounted for 55.4% in all ampicillin-resistant H. influenzae strains. Among them eighteen H. influenzae strains belonged to one type, accounted for 24.3% in all ampicillin-resistant H. influenzae strains. Thirty one genotypes were identified by multiplex PCR assay for ampicillin-resistant H. influenzae. The identity ratio of PFGE and multiplex PCR was 63.5%. CONCLUSION: In Beijing, Shanghai and Guangzhou areas 55.4% of ampicillin-resistant H. influenzae strains had clonal dissemination during the 4 years.
Assuntos
Resistência a Ampicilina/genética , Antibacterianos/farmacologia , Farmacorresistência Bacteriana/genética , Infecções por Haemophilus/epidemiologia , Haemophilus influenzae/genética , Pré-Escolar , China/epidemiologia , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Genótipo , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/classificação , Haemophilus influenzae/isolamento & purificação , Humanos , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Nasofaringe/microbiologia , Reação em Cadeia da Polimerase , Infecções Respiratórias/microbiologiaRESUMO
OBJECTIVE: To analyze the mechanisms of macrolide resistance in Streptococcus pneumoniae from children in Beijing. METHODS: The MICs of penicillin and erythromycin were determined by the E-test methods for 200 Streptococcus pneumoniae isolates collected from 2002 to 2003 at Beijing Children's Hospital. MICs of azithrhomycin, clarithromycin, acetylspiramycin and clindamycin for 147 erythromycin-resistant isolates were detected by the agar dilution methods. For phenotyping, macrolide resistance induction tests were used in erythromycin-resistant isolates. PCR was used to determine the presence of the erythromycin-resistant genes. RESULTS: Of 200 Streptococcus pneumoniae isolates, 89.5% were resistant to erythromycin. In 147 erythromycin-resistant isolates, resistance rates were as follows: azithromycin, 100%; clarithromycin, 100%; acetylspiramycin, 95.2%; and clindamycin, 95.9%. The most common macrolide resistance phenotype was the cMIS phenotype (95.9%), 1.4% had the iMLS phenotype and 2.7% the M phenotype. Erythromycin-resistant isolates were characterized for the underlying resistance genotype, with 79.6% having the ermB genotypes, 17.7% having both ermB and mefA, 2.7% having the mefA, and none having neither ermB nor mefA genotypes. CONCLUSIONS: The rates of carriage of macrolide-resistant Streptococcus pneumoniae by children were high in Beijing during 2002 - 2003. cMLS was the most prevalent phenotype among erythromycin-resistant Streptococcus pneumoniae isolates, and ribosomal modification (ermB gene coded) was the main resistance mechanism against macrolides in Beijing region.