Xuzhuang Shen

The prevalence of plasmid-mediated AmpC β-lactamases among clinical isolates of Escherichia coli and Klebsiella pneumoniae from five children’s hospitals in China

The purpose of this study was to investigate the prevalence of plasmid-mediated AmpC β-lactamases in Escherichia coli and Klebsiella pneumoniae from five children’s hospitals in China. A total of 494 E. coli and 637 K. pneumoniae isolates were collected from five children’s hospitals in China from 2005 to 2006. The isolates with decreased susceptibility to cefoxitin were subjected to confirmation test with 3-aminophenyl boronic acid. Polymerase chain reaction (PCR) amplification of the blaAmpC, blaTEM, blaCTXM, and blaSHV genes and their gene sequencing were performed. Transconjugants were achieved by conjugation experiments. Plasmid-mediated AmpC β-lactamases were found in 10.1% of K. pneumoniae (64/637) and in 2.0% of E. coli (10/494) strains. The proportion of plasmid-mediated AmpC-producing strains significantly increased from 2005 (2.6%) to 2006 (9.3%) (p<0.001). The DHA-1-producing isolates were the most prevalent type (93.2%, 69/74). The sequences of blaDHA-1 genes were all identical to those from the GenBank. Strains of blaCMY-2 were isolated from five isolates (6.8%), which were all from E. coli. One sequence of blaCMY-2 differs from blaCMY-2 in the GenBank. Eighteen of the 74 (24.3%) AmpC-producing K. pneumoniae and E. coli isolates coproduced an extended-spectrum β-lactamase (ESBL). Cefoxitin resistance was transferred to 15 of the 74 positive strains (20.3%). Our study has demonstrated the occurrence of plasmid-mediated AmpC β-lactamases in E. coli and K. pneumoniae in Chinese pediatric patients and DHA-1 type AmpC enzymes had the highest prevalent rate. The CMY-2 AmpC β-lactamases from the children’s hospitals in China in this study are the first reported. Hence, continuous surveillance of the prevalence and evolution of AmpC β-lactamase is important.

Molecular epidemiological analysis of methicillin-resistant Staphylococcus aureus isolates from Chinese pediatric patients.

To investigate the molecular epidemiological analysis of methicillin-resistant Staphylococcus aureus (MRSA) isolates from five pediatric hospitals in China. Seventy-three MRSA isolates were analyzed by a combination of different genotyping methods, including multilocus sequence typing (MLST), SCCmec and spa typing. Panton-Valentine Leukocin (PVL) gene was also detected. The prevalent strains were ST239-MRSA-III and ST1-MRSA clones in the northern region; ST239-MRSA-III, ST910-MRSA-IV and ST88-MRSA in the eastern region; and ST59-MRSA in the southern region. Only the ST910-MRSA-IV clone has been found in China until now, and it is closely related to ST30-MRSA-IV. All MRSA isolates were found to be resistant to penicillin and azithromycin, and multidrug resistance was observed. The cases of necrotic pneumonia, severe skin and subcutaneous tissue infection and lymphadenitis resulted from PVL gene-positive MRSA. There were several novel genetic types of MRSA. Antimicrobial susceptibility tests showed high resistance of many antimicrobials and multiple drugs.

Antibiotic use in five children's hospitals during 2002–2006: the impact of antibiotic guidelines issued by the Chinese Ministry of Health

To investigate the pattern of antibiotic use in five Chinese childrens hospitals from 2002 to 2006. To see if the Guidelines to encourage rational use of antibiotics issued by the Ministry of Health in October 2004 have any impact on the use.

Outpatient antibiotic use and assessment of antibiotic guidelines in Chinese children’s hospitals

PurposeOur aim was to investigate outpatient antibiotic use and assess the impacts of intervention in five Chinese children’s hospitals from 2002 to 2006.MethodsThe Anatomical Therapeutical Chemical Classification/Defined Daily Doses and Drug Utilization 90% methodologies were used. We also analyzed the relationship between antibiotic consumption and resistant rate in one of the hospitals.ResultsThe overall antibiotic consumption decreased during the intervention period in some hospitals, and the variation in use between hospitals was also reduced. A decrease in penicillins and first-generation cephalosporins, and an increase in third-generation cephalosporins as well as in the combinations of penicillins and β-lactamase inhibitors were observed. In addition, an increasing resistance to antibiotics was a concern due to antibiotic overconsumption.ConclusionsThe intervention had effects on the overall antibiotic use in outpatients. However, over the span of 5 years, there was a decrease in the use of narrow-spectrum antibiotics and an increase in broad-spectrum antibiotics.

Occurrence of qnr‐positive clinical isolates in Klebsiella pneumoniae producing ESBL or AmpC‐type β‐lactamase from five pediatric hospitals in China

The plasmid-mediated quinolone resistance qnr genes in clinical isolates in adults have been described in different countries; however, the frequency of their occurrence has not been detected in pediatric patients. A total of 410 clinical isolates of Klebsiella pneumoniae, identified as producers of an extended-spectrum beta-lactamase (ESBL), or AmpC beta-lactamase, were collected from five childrens hospitals in China during 2005-2006. The isolates were screened for the presence of the qnrA, qnrB, and qnrS genes, and then the harboring qnr gene isolates were detected for a bla gene coding for the TEM, SHV, CTX-M, and plasmid-mediated ampC gene by a PCR experiment. Ninety-two isolates (22.7%) were positive for the qnr gene, including 10 of qnrA (2.4%), 25 of qnrB (6.1%), and 62 of qnrS (15.1%). Eighty-one of the 92 (88.0%) qnr-positive isolates carried at least one bla gene for TEM, SHV, CTX-M, or DHA-1. The ciprofloxacin resistance increased 16-256-fold and oflaxacin resistance increased 2-32-fold in transconjugants, respectively. These results indicated that the plasmid-mediated qnr quinolone resistance gene was qnrS, followed by qnrB and qnrA. Most of the isolates also carried a bla gene coding ESBL or ampC gene coding DHA-1 among Klebsiella pneumoniae isolated from Chinese pediatric patients.

Molecular and clinical characteristics of clonal complex 59 methicillin-resistant Staphylococcus aureus infections in Mainland China.

Detailed molecular analyses of Clonal Complex 59 (CC59) methicillin-resistant Staphylococcus aureus (MRSA) isolates from children in seven major cities across Mainland China were examined. A total of 110 CC59 isolates from invasive and non-invasive diseases were analyzed by multilocus sequence typing (MLST), Staphylococcus cassette chromosome mec (SCCmec) typing, staphylococcal protein A (spa) typing and pulsed-field gel electrophoresis (PFGE). Antibiotics susceptibilities, carriage of plasmids and 42 virulence genes and the expression of virulence factors were examined. ST59 (101/110, 91.8%) was the predominant sequence type (ST), while single locus variants (SLVs) belonging to ST338 (8/110, 7.3%) and ST375 (1/110, 0.9%) were obtained. Three SCCmec types were found, namely type III (2.7%), type IV (74.5%) and type V (22.7%). Seven spa types including t437, which accounted for 87.3%, were determined. Thirteen PFGE types were obtained. PFGE types A and B were the major types totally accounting for 81.8%. The dominant clone was ST59-t437-IVa (65.5%), followed by ST59-t437-V (14.5%). The positive rate of luks-PV and lukF-PV PVL encoding (pvl) gene was 55.5%. Plasmids were detected in 83.6% (92/110) of the strains. The plasmid size ranging from 23.4 kb to 50 kb was most prevalent which accounted for 83.7% (77/92). A significantly lower expression of hla was found in ST59-t437-IVa compared with ST59-t437-V. Among the 110 cases, 61.8% of the patients were less than 1 year old. A total of 90 cases (81.8%) were community-associated (CA) infections whereas 20 cases (18.2%) were hospital-associated (HA) infections. Out of the 110 patients, 36.4% (40/110) were diagnosed with invasive infectious diseases in which ST59-t437-IVa accounted for 67.5% (27/40). In brief, ST59-t437-IVa was proved as the dominant clone in CC59 MRSA strains. The carriage rate of pvl gene was high. CC59 MRSA could result in CA and HA infections. The majortiy of MRSA infection children were in young age.

Antimicrobial Resistance and Serotypes of Nasopharyngeal Strains of Streptococcus pneumoniae in Chinese Children with Acute Respiratory Infections

This prospective, multicentre, nasal carriage study in Chinese children with upper respiratory infection was carried out over the period from 2000 to 2002. Overall, the prevalence of pneumococcal carriage was 24.9%. Antimicrobial susceptibility tests were performed for 887 isolates of Streptococcus pneumoniae of which 33.5% were intermediately susceptible to penicillin and 6.4% were resistant. Multidrug resistance was very common. Pneumococcal strains (n = 625) were serotyped, showing 72.2% were covered by the 23-valent pneumococcal polysaccharide vaccine and 57.6% by the seven-valent pneumococcal conjugate vaccine. Sero-groups 19 and 23 were significantly associated with penicillin resistance, which is increasing in China. Erythromycin, tetracycline and sulphamethoxazole/trimethoprim cannot be recommended as first-line treatments for respiratory tract infection as in some other developing countries. These features of serotype distribution are of importance for surveillance in the era of the new conjugate vaccine. In particular, these features will allow for documentation of serotype replacement after the introduction of widespread vaccination.

Characterization of emm types and superantigens of Streptococcus pyogenes isolates from children during two sampling periods.

The characteristics of 359 group A streptococcal (GAS) isolates collected from Chinese paediatric patients in two periods (1993-1994, 2005-2006) were studied. Isolates were assigned to emm types and assayed for eight superantigen (SAg) genes (speA, speC, speH, speI, speG, speJ, ssa, SMEZ). Types emm1 and emm12 were consistently the most prevalent during the two periods, while others varied in frequency. GAS isolates carrying six or more SAg genes increased from 46.53% (1993-1994) to 78.39% (2005-2006); ssa, speH and speJ genes (P<0.05) increased but speA declined (P<0.05). SAg gene profiles were closely associated with the emm type, but strains of the same emm type sometimes carried different SAg genes in the two periods. No significant difference in emm-type distribution and SAg gene profile was noted between isolates from different diseases. These data may contribute towards the development of a GAS vaccine in China.

Drug utilisation 90% (DU90%) profiles of antibiotics in five Chinese children's hospitals (2002-2006).

This study surveyed the pattern of inpatient antibiotic use in five Chinese childrens hospitals between 2002 and 2006, focusing on the antibiotics accounting for 90% of the volume utilised as well as the level of adherence to guidelines. The Anatomical Therapeutical Chemical Classification/Defined Daily Doses (ATC/DDD) and the drug utilisation 90% (DU90%) methodologies were used. In October 2004, national antibiotic guidelines were issued that divided antibiotics into non-restricted, restricted and special use grades. In five childrens hospitals, a total of 56 different systemic antibiotics were used during the study period. Antibiotics that could be injected accounted for 59.0-99.8%. beta-Lactam antibacterials (ATC codes J01C and J01D) were the most used subgroups. The numbers of antimicrobial agents and non-restricted antibiotics within the DU90% segment were 11-20 and 5-9, respectively. The proportion of non-restricted antibiotic consumption was ca. 40% in 2006 in four hospitals, which varied among hospitals during the period 2002-2005. There was considerable variation both in the pattern and amount of antibiotics used in the five hospitals, with amoxicillin/clavulanic acid, cefuroxime and cefazolin being the most commonly used substances. We also observed a decrease in the ranks of some restricted and special use antibiotics after the guidelines were issued in one hospital. The DU90% profiles were proven to be useful in studying the pattern of antibiotic use in hospitals. Finally, the study observed the effectiveness of guidelines for antibiotic use in some hospitals, although injectable antibiotics were widely used in childrens hospitals in China.

Skin and soft tissue infections caused by community‐associated methicillin‐resistant staphylococcus aureus among children in China

Aim:  To investigate the characteristic of community‐associated methicillin‐resistant staphylococcus aureus (CA‐MRSA) skin and soft tissue infections (SSTIs) among children in China.