Molecular characteristics of methicillin resistant Staphylococcus aureus isolated from Chinese children / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) isolates from Chinese children in seven cities.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>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.</p>

Distribution and antibiotic resistance of pathogens isolated from children with infectious diarrhea in Guangzhou / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the distribution and antibiotic resistance of the isolated pathogens from children with infectious diarrhea in Guangzhou.</p><p><b>METHODS</b>The fecal samples of 2 409 children with infectious diarrhea between January 2006 and December 2007 were collected and cultured. Pathogenic bacterium were isolated and identified by biochemical and serological methods. The antibiotic susceptibilities were tested by the Kirby-Bauer method.</p><p><b>RESULTS</b>A total of 448 isolates of pathogenic bacterium (18.6%) were obtained, including Shigella (n=159), enteropathogenic Escherichia coli (n=141), Salmonella (n=76), Vibrion (n=11), fungus (n=41), and C jejuni (n=20). All of isolates of the three major pathogenic bacterium, Shigella, enteropathogenic Escherichia coli and Salmonella, were susceptible to imipenem and less than 10% of the isolates were resistant to the third generation cephalosporins and beta-lactamase inhibitors. However, the isolates showed a high resistance to ampicillin and sulfamethoxazole/trimethoprim (>75%).</p><p><b>CONCLUSIONS</b>Shigella, enteropathogenic Escherichia coli and Salmonella were major pathogenic bacterium of diarrhea in children from Guangzhou. The major isolates were susceptible to imipenem, the third generation cephalosporins and beta -lactamase inhibitors, but were resistant to ampicillin and sulfamethoxazole/trimethoprim.</p>

Cloning and sequence analysis of UreB of Helicobacter pylori isolated from children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To clone UreB gene of Helicobacter pylori (H. pylori) isolated from children to pGEX-4T-1 expression plasmid, and do sequence analysis.</p><p><b>METHODS</b>A pair of specific primer was designed according to H. pylori UreB gene in the GenBank. Using H. pylori strains isolated from children as a template, a UreB gene was obtained by PCR. After EcoR I and Not I digestion, the PCR production was linked with pGEX-4T-1 which was digested with the same enzymes. The recombinant plasmid was transformed into E.coli BL21 and identified by double enzyme digestion and sequence analysis. The sequence results were compared with the gene sequence in the GenBank.</p><p><b>RESULTS</b>A UreB gene was successfully amplified from children's H. pylori strain GZCH1. It was 1710 bp in size. The objective band was identified by double enzyme digestion. DNA sequence showed that UreB was in the correct open reading frame. The sequence comparison analysis showed that DNA and amino acid sequence identities of UreB gene with other strains were 98%. The sequence of UreB of H. pylori strain GZCH1 was submitted to GenBank (accession number:FJ455126).</p><p><b>CONCLUSIONS</b>UreB of H. pylori strain GZCH1 is successfully cloned to pGEX-4T-1, which provides a basis for research of oral H. pylori vaccine.</p>

Investigation of campylobacter jejuni infection in children with diarrhea in Guangzhou / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the incidence of campylobacter jejuni (CJ) infection and the drug resistance of CJ in children with diarrhea in Guangzhou.</p><p><b>METHODS</b>The fecal samples of 3,351 children with diarrhea between July 2005 and June 2008 were collected for CJ culture. The species of CJ strains were identified by Lior methods. The drug susceptibility tests were performed by the Kirby-Bauer method.</p><p><b>RESULTS</b>Two hundred and sixty-seven CJ strains (8.0%) were isolated from 3,351 samples. The children at age of 1 month to 1 year were susceptible to CJ, accounting for 91.0%. A higher incidence of CJ infection (76.8%) was found in summer and autumn. The CJ strains were susceptible to imipenem, amikacin, cefoperazone/sulbactam, chloramphenicol, macrolides and lincomycins. Parts of CJ strains (20%-40%) were resistant to ampicillin, quinolones and ambramycin. All CJ strains were resistant to sulfamethoxazole/trimethoprim and cefditoren. Two hundred and one strains (75.3%) were CJ biotype I.</p><p><b>CONCLUSIONS</b>CJ is an important pathogen of diarrhea in children from Guangzhou. CJ is resistant to some antibiotics used often in clinical practice, and so it is thus important to use antibiotics based on the results of drug susceptibility tests in children with CJ infection.</p>

Antimicrobial resistance and penicillin resistance-associated genes of Streptococcus pneumoniae isolated from children with respiratory tract infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the antimicrobial resistance and penicillin resistance-associated genes (TEM and pbp2B) of Streptococcus pneumoniae (S. pneumoniae) isolated from sputum specimens of Guangzhou children with respiratory tract infection.</p><p><b>METHODS</b>E-test and Kirby-Bauer methods were applied to detect the antibiotic susceptibility of 44 strains of S. pneumoniae. PCR was used to detect resistance genes pbp2B and TEM, followed by DNA sequence analysis of pbp2B gene. The sequence results were compared to those of penicillin-susceptible S. pneumoniae R6.</p><p><b>RESULTS</b>Of the 44 isolates of S. pneumoniae, only 5 (11.4%) were susceptible to penicillin. All strains were resistant to erythromycin but susceptible to ofloxacin and vancomycin. The resistance rate of the isolates to clindamycin and trimoxazole was more than 90%. The S. pneumoniae isolates showed a high susceptibility to amoxicillin, imipenem and ceftriaxone, with a resistance rate of 0, 2.6% and 3.9%, respectively. The sequence analysis showed that more than 99% nucleotide sequence of pbp2B gene of five penicillin-susceptible isolates was the same as penicillin-susceptible S. pneumoniae R6, without any amino acid replacement. Site mutation was found in the remaining 39 penicillin-nonsusceptible isolates with a nucleotide mutation rate ranging from 13.2% to 23.1% and amino acid replacement rate from 6.5% to 10.9%. The 39 penicillin-nonsusceptible isolates were classified into 4 types according to the mutation site between Ser391 and Thr492 of pbp2B: type I (n=30), type II (n=7), type III (n=1) and type IV (n=1). No TEM gene was detected in all the 44 S. pneumoniae isolates.</p><p><b>CONCLUSIONS</b>The S.pneumoniae isolates from Guangzhou children with respiratory tract infection are resistant to penicillin and erythromycin. Amoxicillin and the third generation cephalosporin may be recommended for treating S. pneumoniae infection. The mutation of pbp2B gene plays an important role in the development of S. pneumoniae resistance to penicillin.</p>

Surveillance of antibiotic resistance of Streptococcus pneumoniae isolated from hospitalized patients with pneumonia in four children's hospitals in China / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the antimicrobial resistance of Streptococcus pneumoniae (S.pneumoniae) isolated from Chinese children with pneumonia.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Serotype distribution and resistance to beta-lactams of Streptococcus pneumoniae isolated from children in Beijing, Shanghai and Guangzhou, 2000 - 2002 / 中华儿科杂志

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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).</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>

Surveillance of antimicrobial susceptibility of Streptococcus pneumoniae and Hemophilus influenzae isolated from children in Guangzhou area between 2003 and 2004 / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the situation of antibiotic resistance of Streptococcus pneumoniae (Sp) and Hemophilus influenzae (Hi) clinical isolates from children in Guangzhou area.</p><p><b>METHODS</b>The authors cultured, isolated and identified the Sp and Hi strains from nasopharyngeal secretion of patients who visited Guangzhou Children's Hospital for upper respiratory tract infection between 2003 and 2004. K-B disc diffusion and E-test for antibiotic susceptibility were performed for these clinical isolates.</p><p><b>RESULTS</b>Totally 172 and 484 strains of Sp and Hi were respectively isolated from nasopharyngeal secretions in the hospital. For Sp strains, the rates of resistance to penicillin, amoxicillin/clavulanic acid, ceftriaxone, cefuroxime, cefaclor, erythromycin, tetracycline, chloramphenicol, sulfamethoxazole/trimethoprim (SMZ/TMP), clindamycin and ofloxacin were 32.0%, 11.1%, 32.6%, 18.1%, 39.5%, 82.6%, 78.5%, 24.4%, 87.2%, 69.2% and 3.1%, respectively. The penicillin non-susceptible Sp (PNSSP) isolates showed higher rates of resistance to other antimicrobial agents such as other beta-lactam antimicrobial agents, erythromycin, and SMZ/TMP than those of penicillin susceptible Sp (PSSP) isolates. More than 90% of PNSSP were multidrug resistant strains. The average rate of beta-lactamase production among 484 strains of Hi was 29.5% (143/484). For Hi isolates, the rates of resistance to ampicillin, amoxicillin/clavulanic acid, ceftriaxone, cefuroxime, cefaclor, SMZ/TMP, tetracycline, chloramphenicol, azithromycin, and ofloxacin were 40.1%, 3.4%, 4.1%, 1.9%, 5.6%, 56.2%, 52.1%, 17.4%, 2.1%, and 0.6%, respectively.</p><p><b>CONCLUSION</b>The antimicrobials resistant Sp and Hi isolated from children with respiratory tract infection in the area have become a severe problem. The rate of resistance to penicillin of Sp had been decreased compared with the last three years, but the rate of resistance to ceftriaxone of Sp increased, and the multidrug resistance rates of PNSSP was rather high. PNSSP was characterized by a multidrug-resistance to erythromycin, tetracycline and SMZ/TMP. beta-lactamase production and ampicillin resistance among the Hi isolates from children in the area had increased generally during the period 2003 - 2004. The Hi isolates were more susceptible to the second and the third generation cephalosporins, amoxicillin/clavulanic acid and azithromycin.</p>