[Show abstract][Hide abstract] ABSTRACT: Methicillin-resistant Staphylococcus aureus (MRSA) has become an important nosocomial pathogen, causing considerable morbidity and mortality. During the last 20 years, a variety of genotyping methods have been introduced for screening the prevalence of MRSA. In this study, we developed and evaluated an improved approach capillary gel electrophoresis based multilocus variable-number tandem-repeat fingerprinting (CGE/MLVF) for rapid MRSA typing. A total of 42 well-characterized strains and 116 non-repetitive clinical MRSA isolates collected from six hospitals in northeast China between 2009 and 2010 were tested. The results obtained by CGE/MLVF against clinical isolates were compared with traditional MLVF, spa typing, Multilocus sequence typing/ staphylococcal cassette chromosome mec (MLST/SCCmec) and pulse field gel electrophoresis (PFGE). The discriminatory power estimated by Simpson's index of diversity was 0.855 (28 types), 0.855 (28 patterns), 0.623 (11 types), 0.517 (8 types) and 0.854 (28 patterns) for CGE/MLVF, traditional MLVF, spa typing, MLST/SCCmec and PFGE, respectively. All methods tested showed a satisfied concordance in clonal complex level calculated by adjusted Rand's coefficient. CGE/MLVF showed better reproducibility and accuracy than traditional MLVF and PFGE methods. In addition, the CGE/MLVF has potential to produce portable results. In conclusion, CGE/MLVF is a rapid and easy to use MRSA typing method with lower cost, good reproducibility and high discriminatory power for monitoring the outbreak and clonal spread of MRSA isolates.
Full-text · Article · Jan 2014 · International Journal of Molecular Sciences
[Show abstract][Hide abstract] ABSTRACT: To investigate the drug-resistance rates of community-acquired respiratory tract pathogens isolated from adults in China during 2009 and 2010.
A total of 1793 strains (S. aureus 421, S. pneumoniae 420, K. pneumoniae 404, H. influenzae 313, other Streptococcus. spp 149, and M. catarrhalis 86) of non-duplicated community-acquired respiratory tract pathogens were isolated from 11 hospitals in 6 cities. The MIC values were determined by the broth microdilution method, and the production of β-lactamase was tested using a nitrocefin-based test.
All of the S.aureus isolates were methicillin-sensitive (MSSA). Of the MSSA isolates, less than 1% (4/421) was resistant to β-lactamase inhibitor combinations, about 13.1% (55/421) and 9% (38/421) resistant to levofloxacin and moxifloxacin, and 57% (240/421), 53.2% (224/421), and 88.7% (373/421) resistant to azithromycin, clarithromycin, and penicillin, respectively. No S. aureus isolates resistant to vancomycin were detected in this study. Based on different criteria, the percentages of penicillin-sensitive S. pneumoniae (PSSP), penicillin-intermediate S. pneumoniae (PISP), and penicillin-resistant S. pneumoniae (PRSP) were 24.4% (102/420), 27.3% (115/420), 48.3% (203/420) (Oral) and 1.9% (8/420), 9% (38/420), 89.1% (374/420) (parenteral), respectively. The resistance rates of S. pneumonia to azithromycin, clarithromycin, cefaclor, cefuroxime, ceftriaxone and amoxicillin with clavulanic acid were 88.2% (370/420), 87.4% (367/420), 45.3% (190/420), 41.9% (176/420), 10.2% (43/420), and 5.2% (22/420), respectively. About 2.6% (11/420) and 0.2% (1/420) of S. pneumonia isolates were resistant to levofloxacin and moxifloxacin. More than 70% (104/149) of β-hemelutic streptococci isolates were resistant to azithromycin and clarithromycin, and about 10.1% (15/149) of isolates were resistant to levofloxacin. The resistance rates of K.pneumonia to most antibiotics were > 20% (81/404), and that of ceftazidime was lower than cefuroxime, cefaclor, and ceftriaxone. The mean prevalence value of ESBL producing K. pneumonia was 38.8% (157/404), with significantly regional variations. More than 90% of H. influenza and M. catarrhalis were susceptible to most antibiotics, with resistance rate of < 5% (16/313, H. influenza; 4/86, M. catarrhalis). The mean productions of β-lactamase in H. influenza and M. catarrhalis were 13.1% (41/313) and 91.7% (79/86), respectively.
The percentage of PRSP increased significantly, and the resistance rates of community-acquired respiratory tract pathogens to common antibiotics such as macrolide and cephalosporins increased gradually. New fluoroquinolones such as moxifloxacin showed a high antimicrobial activity against most of the respiratory pathogens.
No preview · Article · Feb 2012 · Zhonghua jie he he hu xi za zhi = Zhonghua jiehe he huxi zazhi = Chinese journal of tuberculosis and respiratory diseases
[Show abstract][Hide abstract] ABSTRACT: To investigate antimicrobial resistance among gram-positive cocci in China in 2009.
From June to December 2009, 1169 consecutive and non-repetitive gram-positive cocci were collected from 12 teaching hospitals at 9 cities. The minimal inhibitory concentration (MIC) of antibacterial agents was determined by agar dilution method.
The prevalences of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative Staphylococci (MRCoNS) were 45.3% (211/466) and 89.5% (214/239), respectively. The isolation rate of MRSA was 33.3% - 68.1% from different samples. All Staphylococci isolates were susceptible to vancomycin, teicoplanin and linezolid. Five point five percent (7/128) E.faecium strains were resistant to vancomycin. All E. faecalis strains were susceptible to vancomycin. About 99.1% (108/109) of E.faecalis and E.faecium were susceptible to linezolid. The prevalence of penicillin-intermediate Streptococcus pneumoniae (PISP) was 21.6% (48/222). Only 1 (0.5%, 1/222) Streptococcus pneumoniae strain was resistant to penicillin. Teicoplanin, vancomycin, linezolid and tigecycline were the most active agents against Streptococcus pneumoniae (susceptible rate 100%).
The high prevalence of methicillin-resistance is among Staphylococcus strains. Different samples show a different MRSA prevalence. Teicoplanin, vancomycin and linezolid show very high activity to Staphylococci, E. faecalis, E. faecium and Streptococcus pneumoniae.
No preview · Article · Sep 2010 · Zhonghua nei ke za zhi [Chinese journal of internal medicine]
[Show abstract][Hide abstract] ABSTRACT: To investigate the source and genetic background of methicillin-resistant Staphylococcus aureus in the year of 2006, in China.
From January to December 2006, a total number of 302 consecutive and non-repetitive methicillin-resistant Staphylococcus aureus were collected from 17 Teaching hospitals in 15 areas. Genotypes of SCCmec were determined by multiplex PCR and multilocus sequence typing (MLST) was used to type the house-keeping genes. The implementation of the spa typing method was straightforward, and the results obtained were reproducible, unambiguous, and easily interpreted.
All areas but Dalian harbored SCCmec III while Dalian harbored SCCmec II most. There were two strains in Guangzhou, harboring SCCmec IV. There were four strains of sequence type (ST), with ST239 accounted for 46.7% and ST5 accounted for 44.4%. ST59 accounted for 6.7% and ST88 accounted for 2.2%. There were fourteen strains of Spa typing, with t30 accounted for 52.6%; t37 accounted for 27.2%; t2 accounted for 12.9%; t632 accounted for 2.3%; t437 accounted for 1.3%; t570, t601 accounted for 0.7%; t377, t459, t796, t899, t1152, t2649 accounted for 0.3%; no-typing accounted for 0.3%, respectively. pvl gene was not detected.
The main clone strains were ST239-MRSA-SCCmec III-t30, ST5-MRSA-SCCmec II-t2, with unique geographic distributions across the whole nation.
No preview · Article · Mar 2010 · Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi
[Show abstract][Hide abstract] ABSTRACT: To investigate the antimicrobial resistance of community respiratory pathogens isolated in China.
The strains of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, S. pyogenes were isolated from patients with community-acquired respiratory tract infections at 14 Chinese hospitals from 2005 to 2007. Etest and disk diffusion methods were used to survey the susceptibility of 14 antibiotics against these strains. These antibiotics included penicillin G, ampicillin, amoxicillin/clavulanic acid, cefaclor, cefprozil, ceftriaxone, cefepime, levofloxacin, gatifloxacin, ciprofloxacin, tetracycline, clindamycin, erythromycin and trimethoprim/sulfamethoxazole (SXT).
A total of 1870 strains were collected including S. pneumoniae (n = 997), S. pyogenes (n = 176), H. influenzae (n = 499) and M. catarrhalis (n = 198). The 2005 - 2007 prevalence of penicillin-susceptible S. pneumoniae (PSSP) were 92.6%, 73.9%, 74.1% and penicillin-intermediate S. pneumoniae (PISP) 4.5%, 9.5%, 14.3% and penicillin-resistant S. pneumoniae (PRSP) 2.9%, 16.6%, 11.6% respectively. 36.9% of S. pneumoniae strains isolated from <or= 6 years old children were penicillin-non-susceptive isolates (PNSSP) and < 22.0% of PNSSP isolated from other age groups. The susceptible rates of beta-lactamase antibiotics to PRSP and PISP isolates were less than 25.0% and 49.2% respectively. From 48.5% to 98.6% PSSP isolates were susceptible to beta-lactamase antibiotics. The susceptible rates of PNSSP and PSSP to erythromycin, tetracycline and SXT were below 7.1% and 32.1% respectively. About 95% S. pneumoniae were susceptible to ciprofloxacin, levofloxacin and gatifloxacin. All of S. pyogenes isolates were susceptible to beta-lactamase antibiotics, and 16.7%, 27.1% and 15.6% Of S. pyogenes isolates were susceptible to erythromycin. 8.5%, 19.9%, 15.3% of H. influenzae and 57.4%, 78.8%, 95.5% of M. catarrhalis produced beta-lactamase during the 3-year period. The susceptible rates of cefepime, ceftriaxone, gatifloxacin, levofloxacin and ciprofloxacin to H. influenzae and M. catarrhalis were >or= 92.9%.
Antimicrobial resistance in S. pneumoniae is rising. The prevalence of PNSSP isolated from children < or = 6 years old is higher than other age groups. Amoxicillin-clavulanic acid, ceftriaxone, cefepime, gatifloxacin and levofloxacin remain highly active against common community respiratory pathogens.
No preview · Article · Nov 2009 · Zhonghua yi xue za zhi
[Show abstract][Hide abstract] ABSTRACT: To evaluate the in vitro activity of daptomycin, vancomycin, teicoplanin, tigecycline, ceftobiprole and linezolid against 499 strains of blood-isolated gram-positive cocci.
Determination of the minimal inhibitory concentration (MICs) of daptomycin with microbrothdilution method and the MICs of other 9 antimicrobial agents with agar dilution method against 499 strains of blood-isolated gram positive cocci was carried out. The data was analyzed with WHONET 5.4 software.
The susceptibility rates of staphylococci to daptomycin, tigecycline, linezolid, ceftobiprole, vancomycin and teicoplanin were 100%. All staphylococcus strains were inhibited by daptomycin at a MIC of 1 mg/L. The MIC(50) and MIC(90) of daptomycin were both 0.5 mg/L against methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant Staphylococcus coagulase-negative (MRSCoN). Among Enterococcus spp, the highest MIC of daptomycin was 4 mg/L. The MIC(50) and MIC(90) of daptomycin were both 2 mg/L against E.faecalis, whereas they were 2 mg/L and 4 mg/L against E.faecium. One strains of linezolid-resistant E.faecalis (MIC: 8 mg/L) was susceptible to daptomycin (MIC: 1 mg/L). Three strains of E.faecium carrying vanA gene with vancomycin MICs above 32 mg/L and teicoplanin MICs also 32 mg/L were susceptible to daptomycin, tigecycline and linezolid. The MIC range of daptomycin against Streptococcus pneumoniae and Streptococcus viridans was 0.032 - 0.25 mg/L and 0.125 - 1.000 mg/L separately.
Daptomycin has excellent in vitro activity against common gram-positive pathogens isolated from blood. It may be a good choice for clinicians to treat drug-resistant gram-positive cocci.
No preview · Article · Apr 2009 · Zhonghua nei ke za zhi [Chinese journal of internal medicine]
[Show abstract][Hide abstract] ABSTRACT: To investigate the clinical characteristics of methicillin-resistant Staphylococcus aureus (MRSA) in phlegm specimens of positive patients, so as to provide evidences for the nosocomial infection control.
We retrospectively analyzed the clinical data of 211 hospitalized patients who were MRSA-positive in their phlegm specimens in PUMC Hospital from January 2005 to October 2007.
Among the 211 patients, 196 (92.9%) had received antibiotics three months before the detection of MRSA, and 128 (60.7%) had received more than three antibiotics. Over 90% of MRSA were resistant to levofloxacin, erythromycin, clindamycin, and gentamicin, and 73.9% were resistant to rifampicin.
Improper use of antibiotics should be avoided. Vancomycin is the first choice for MRSA treatment.
No preview · Article · Nov 2008 · Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae
[Show abstract][Hide abstract] ABSTRACT: To investigate and take a case study on a Klebsiella pneumoniae outbreak in a newborn intensive care unit (NICU).
Using epidemiological investigation method to cultivate bacilli and detect the homology.
Klebsiella pneumonia was detected in 4 NICU patients. Based on environmental sample analyses, four Klebsiella pneumonia strains were identified and confirmed to be highly homologous. The outbreak was effectively controlled after the strict implementation of hand hygiene practice and environment disinfection.
Klebsiella pneumonia outbreak in NICU may be caused by the route of hand transmission.
No preview · Article · Nov 2008 · Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae
[Show abstract][Hide abstract] ABSTRACT: To investigate the molecular mechanism of multiple-drug and pan-drug resistance among Acinetobacter species.
Non-repetitive 90 carbapenem-resistant strains of Acinetobacter species were collected in Beijing, Guangzhou, and Fuzhou 1999-2004. The homology of the isolates was determined by both pulsed field gel electrophoresis and randomly amplified polymorphic DNA typing. Seven representative clones were selected from the 90 strains of Acinetobacter isolated from different hospitals to be used for further study. Analytical isoelectric focusing was used to measure the isoelectric point of the beta-lactamase. Plasmid DNA was extracted and purified Genes of different beta-lactamase, including bla(TEM--), bla(SHV-), bla(PER-), blaI(MP-), bla(VIM-), and bla(OXA-) genes, in these clone strains were amplified and sequenced. PCR was used to analyze the integrons.
The P clone strain isolated during an outbreak of pan-drug-resistant Acinetobacter species in Peking Union Medical College Hospital 2004 was not susceptible to most common antimicrobial agents tested. The 7 representative clones produced multiple beta-lactamases: TEM-1, high-level AmpC, SHV-type, OXA-23 carbapenemase and IMP-8 and metalloenzyme respectively. One clone produced PER-1 enzyme. These 7 clone strains were resistant to most beta-lactams (including carbapenems), erythromycin, chloramphenicol, and rifampin. Two clone strains were susceptible to cefoperazone/sulbactam and amikacin while 4 clone strains susceptible to levofloxacin. All of the 7 clones were susceptible to minocycline and colistin. Five different integrons were found, harboring the genes mediating the resistance to aminoglycosides, rifampin, chloramphenicol, and carbapenems (bla(IMP-8)).
The molecular bases of multiple-drug or pan-drug resistance in Acinetobacter species include production of OXA-23 carbapenemase or IMP type metalloenzyme and integrons with different resistance gene cassettes. Pan-drug-resistant Acinetobacter species are susceptible to old antimicrobials agents, such as colistin and minocycline.
No preview · Article · Feb 2006 · Zhonghua yi xue za zhi
[Show abstract][Hide abstract] ABSTRACT: To investigate the prevalence and genotype of plasmid-mediated cephalosporinase (AmpC) beta-lactamase in extended-spectrum-beta-lactamase-producing (ESBL) Escherichia coli and Klebsiella pneumoniae.
24 strains of cefoxitin-resistant ESBL-producing E. coli and 8 strains of K. pneumoniae were collected from January to December 2001 at Beijing Chaoyang Hospital. Analytical isoelectric focusing electrophoresis was used to measure the pI of the beta-lactamase. Conjugation experiment was used to study the transfer of cefoxitin resistance. The homology of the isolates was determined by pulsed field gel electrophoresis (PFGE). Plasmid-mediated AmpC enzyme genes were amplified and sequenced by using multiplex PCR.
The prevalence of ESBL-producing E. coli and K. pneumoniae were 16.8% (49/292) and 16.5% (35/212), respectively. The prevalence of AmpC enzyme among ESBL-producing E. coli and K. pneumoniae isolates were 2.0% (1/49) and 17.1% (6/35), respectively. These 7 isolates produced DHA-1 AmpC enzyme. One strain of K. pneumoniae could transfer cefoxitin resistance to the recipient. Among 7strains, 5 strains produced CTX-M-3 and 2 produced SHV-12 ESBL. These 7 strains also produced TEM-1 broad-spectrum enzymes. These strains harbored 2 - 5 plasmids and one of them were 33 - 36 kb. PFGE showed these strains came from a variety of clones.
In this hospital, 7 strains of the ESBL-positive E. coli and K. pneumoniae produced both DHA-1AmpC enzyme and CTX-M-3/SHV-12 ESBL. These 7 strains were from different clones.
No preview · Article · Aug 2004 · Zhonghua nei ke za zhi [Chinese journal of internal medicine]
[Show abstract][Hide abstract] ABSTRACT: To investigate the mechanism of carbapenems resistance in Acinetobacter baumannii.
WHONET-5 software was used to analyze the trend of carbapenem resistance in Acinetobacter baumannii collected from 1999 to 2001 at Peking Union Medical College Hospital. Analytical isoelectric focusing was used to measure the pI of the beta-lactamase. Conjugation experiment was used to study the transfer of carbapenem resistance and plasmid DNA was extracted and purified with Qiagen Plasmid Mini Kit. The homology of the isolates was determined by pulsed field gel electrophoresis (PFGE). Integrase genes and blaIMP-, blaVIM-, blaOXA- genes for resistant isolates were amplified and sequenced.
Imipenem resistance in A. baumannii was ranged from 1.8%-8.5%, but only 9 resistant isolates were viable. They were co-resistant to other carbapenems, ceftazidime, aztreonam, and gentamicin, and four isolates were resistant to ciprofloxacin. Impipenem resistance could not be transferred to susceptible strains. No plasmid was extracted. Each isolate produced TEM-1, AmpC, and two enzymes (pI 6.7, 6.0), which can not be inhibited by cloxacillin and clavulanic acid. Each isolate had class I intergase gene. Nine isolates were all negative for PCR of blaIMP- and blaVIM- genes, but positive for blaOXA-23 specific PCR. Sequencing found 100% homology with blaOXA-23. PFGE found 3 clones (A type: 5 isolates; B type: 3 isolates; C type: 1 isolate). Control isolates (imipenem-susceptible, but ceftazidime, ciprofloxacin, and gentamicin resistant) were also A clone.
Production of OXA-23 carbapenemase in A. baumannii was one of the main mechanisms of carbapenems resistance at our hospital. It brings concern that imipenem-resistant clone has evoluted from nosocomial multiple-resistant strains.
No preview · Article · Nov 2003 · Zhongguo yi xue ke xue yuan xue bao. Acta Academiae Medicinae Sinicae