Chuanqing Wang

Serotype Distribution and Antimicrobial Resistance of Streptococcus pneumoniae Isolates That Cause Invasive Disease among Chinese Children

A total of 171 Streptococcus pneumoniae isolates causing invasive disease were isolated from Chinese children. The serotype distribution and antimicrobial resistance were tested. The results suggested that the 7-valent pneumococcal conjugate vaccine has a preventive effect among children and that there should be long-term surveillance for serotype 19A.

Superantigen gene profiles and presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus isolated from Chinese children.

This study aimed to evaluate the distribution of superantigen gene profiles and the presence of exfoliative toxin genes in community-acquired meticillin-resistant Staphylococcus aureus (CA-MRSA) isolated from Chinese children, and simultaneously to assess virulence gene profiles and genetic background. Of the CA-MRSA isolates, 88.9 % (88/99) harboured toxin genes, with sek as the most frequent toxin gene (62.6  %), followed by seq (61.6  %), seb (60.6  %) and sea (35.4  %). The eta gene was detected only in one ST398-IVa-spa t034 strain. The sed and etd genes were not found in any of the isolates tested. A total of 38 virulence genotypes were observed, of which the genotype seb-sek-seq (27.3  %, 24/88) comprised the majority, followed by sea-seb-sek-seq (18.2  %, 16/88). The enterotoxin gene cluster including seg-sei-sem-sen-seo-seu predominated at a rate of 15.1  %. The relationship among toxin genotypes, toxin genes encoding profiles of mobile genetic elements and genetic background was analysed. Among 66 clonal complex (CC) 59 isolates, 87.9 % (58/66) were positive for toxin genes, and 75.8  % (50/66) harboured the toxin gene combination seb-sek-seq. Among seb-sek-seq-positive CC59 strains, 42.0  % (21/50) also carried the sea gene. CC59 corresponded exclusively to accessory gene regulator 1 (agr-1). The data presented here enhance our current knowledge on the virulence determinants of CA-MRSA.

Community-acquired, methicillin-resistant Staphylococcus aureus isolated from children with community-onset pneumonia in China.

Community‐acquired, methicillin‐resistant Staphylococcus aureus (CA‐MRSA) has been associated with morbidity and mortality in various countries. In this study, we characterized the molecular and clinical features of pediatric CA‐MRSA pneumonia in China. Between June 2006 and February 2008, 55 previously healthy children confined in eight hospitals countrywide were found to be afflicted with CA‐MRSA pneumonia. A total of 55 strains collected from these children were analyzed by multilocus sequence typing (MLST), Staphylococcus cassette chromosome mec (SCCmec) typing, and spa typing. The Panton–Valentine leukocidin (PVL) gene was also detected. Overall, nine STs were obtained, with ST59 (40.4%) established to be the most prevalent type. We first registered the new ST1409 from a child with necrotizing pneumonia. SCCmecIVa was the most predominant type, followed by SCCmec type V. Twelve spa types were identified, of which one new spa type, t5348, was first detected and registered. One typical livestock‐associated spa type, t034, was found in a 4‐month‐old girl living in the countryside. We also found that 40% of those isolates were PVL‐positive. In addition, the median age of the children in this study was 10 months. A total of 69% (38/55) of the children with community‐acquired pneumonia (CAP) had preceding influenza or influenza‐like illness, and three ST910‐MRSA‐IV strains (PVL gene‐positive) were associated with severe necrosis. The results indicated that the recent CA‐MRSA found in Chinese children with CAP was largely associated with the spread of the ST59‐MRSA‐IV clone, and most of the PVL‐positive strains in this study did not cause necrotic cases. Pediatr Pulmonol. 2010; 45:387–394.

Multidrug-resistant clones of community-associated meticillin-resistant Staphylococcus aureus isolated from Chinese children and the resistance genes to clindamycin and mupirocin

This study aimed to correlate the multidrug resistance (MDR) and sequence type (ST) clones of community-associated (CA) meticillin-resistant Staphylococcus aureus (MRSA) to identify the genes responsible for clindamycin and mupirocin resistance in S. aureus isolates from paediatric hospitals in mainland China. A total of 435 S. aureus isolates were collected. Compared with CA meticillin-susceptible S. aureus (MSSA), the resistance rates of CA-MRSA to ciprofloxacin, chloramphenicol, gentamicin and tetracycline were higher (19.0 vs 2.6 %, P<0.001; 14.7 vs 3.1 %, P<0.001; 14.7 vs 3.1 %, P<0.01; and 46.0 vs 13.3 %, P<0.001, respectively). Compared with hospital-associated (HA)-MRSA, the resistance rates of CA-MRSA to ciprofloxacin, gentamicin, rifampicin, tetracycline and trimethoprim-sulfamethoxazole were lower (19 vs 94.8 %, P<0.001; 14.7 vs 84.4 %, P<0.001; 5.5 vs 88.3 %, P<0.001; 46 vs 94.8 %, P<0.001; and 1.8 vs 9.1 %, P<0.01, respectively). The resistance rates of CA-MRSA, HA-MRSA and CA-MSSA to clindamycin (92.0, 77.9 and 64.1 %, respectively) and erythromycin (85.9, 77.9 and 63.1 %, respectively) were high. The MDR rates (resistance to three or more non-β-lactams) were 49.6, 100 and 14 % in the CA-MRSA, HA-MRSA and CA-MSSA isolates, respectively. Five of seven ST clones in the CA-MRSA isolates, namely ST59, ST338, ST45, ST910 and ST965, had MDR rates of >50 % (67.9, 87.5, 100, 50 and 83.3 %, respectively). The constitutive phenotype of macrolide-lincosamide-streptogramin B (MLS(B)) resistance (69 %) and the ermB gene (38.1 %) predominated among the MLS(B)-resistant CA S. aureus strains. The resistance rate to mupirocin was 2.3 % and plasmids carrying the mupA gene varied in size between 23 and 54.2 kb in six strains with high-level resistance as determined by Southern blot analysis. The present study showed that resistance to non-β-lactams, especially to clindamycin, is high in CA-MRSA isolates from Chinese children and that the profile of resistance is related to clonal type. This study revealed distinctive patterns of MLS(B)-resistant genes among CA S. aureus isolates.

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.

Diversity of pneumococcal surface protein A (PspA) and relation to sequence typing in Streptococcus pneumoniae causing invasive disease in Chinese children

The objective of this paper was to investigate the sequence types (STs) and diversity of surface antigen pneumococcal surface protein A (PspA) in 171 invasive Streptococcus pneumoniae isolates from Chinese children. A total of 171 pneumococci isolates were isolated from Chinese children with invasive pneumococcal diseases (IPD) in 11 hospitals between 2006 and 2008. The pneumococci samples were characterized by serotyping, PspA classification, and multilocus sequence typing (MLST). The PspA of these strains could be assigned to two families. The PspA family 2 was the most common (120/171, 70.1%). No PspA family 3 isolates were detected. Family 1 could be subdivided into two clades, with 42 strains in clade 1 and 9 strains in clade 2, and family 2 could be subdivided into clades 3, 4, and 5, which respectively contained 5, 21, and 14 strains. In total, 65 STs were identified, of which ST320 (30/171, 17.5%), ST271 (23/171, 13.5%), and ST876 (18/171, 10.5%) were the most common types. PspA family 2 and family 1 were dominant among pneumococcal clones isolated from Chinese children with invasive disease. The strains with the same ST always presented in the same PspA family.

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.

Panton–Valentine leukocidin gene sequence variation and phage in methicillin-resistant and methicillin-susceptible Staphylococcus aureus from children in mainland China

To determine the variation in the Panton–Valentine leukocidin (PVL) gene sequences and different PVL‐encoding phages of Staphylococcus aureus strains collected from children in mainland China, fifty‐eight strains with PVL collected from 2007 to 2009 were used. Their molecular characteristics were examined. Primers were designed to sequence the PVL genes. Six PVL‐encoding phages (ϕPVL, ϕ108PVL, ϕSLT, ϕSa2MW, ϕSa2958, and ϕSa2USA) were identified by PCR. Eleven sequence types (ST) were detected with ST59 (39.7%, 23/58) the most frequent ST, followed by 910 (22.4%, 13/58), and 338 (12.1%, 7/58). Single nucleotide polymorphisms (SNP) were identified at 11 locations in the PVL genes. SNP (nucleotide 1396, A→G) and SNP (nucleotide 1546, A→G) were observed in >10 sequences. Four additional SNP were non‐synonymous. Both SNP (nucleotide 16, C→A) and SNP (nucleotide 62, C→T) were present in the same ST59 strain. SNP (nucleotide 527, A→G) was present in five strains belonging to ST30, 121, 1, and 93. SNP (nucleotide 1436, A→C) was present in one ST30 strain. Fifteen strains belonging to ST910, ST217, and ST30 carried a PVL phage that had an icosahedral head morphology. Nine ST59 strains carried ϕ108PVL. Three ST88 strains carried a PVL phage that had an elongated head morphology. Twenty‐seven strains, including 60.9% (14/23) of ST59 and all ST338 strains, had no detectable phage. In conclusion, sequence variation in PVL genes and PVL‐encoding phages was generally related to the lineage. ST59 strains may indeed carry novel PVL phages.

Susceptibility to and resistance determinants of fusidic acid in Staphylococcus aureus isolated from Chinese children with skin and soft tissue infections.

This study aims to determine the resistance rates and determinants of fusidic acid among Staphylococcus aureus isolates collected from Chinese pediatric patients with skin and soft tissue infections (SSTIs). Between 2008 and 2009, a total of 186 clinical S. aureus isolates were collected from the pediatric patients with SSTIs, abscess (44.6%) was the most common SSTI in children 0-16 years old. Four clinical isolates (4/186, 2.2%) were resistant to fusidic acid. Two of these isolates were methicillin-resistant S. aureus (MRSA) that carry the fusC gene. The other two isolates were methicillin-sensitive S. aureus (MSSA) that carry the fusB gene. In the two fusB-positive clinical isolates, the fusB gene was located in a transposon-like element that has 99% identity with a pUB101 fragment from S. aureus. The four fusidic acid-resistant clinical isolates were ST1-MRSA-SCCmecV-t127, ST93-MRSA-SCCmecIII-t202, ST680-MSSA-t5415, and ST680-MSSA-t377. The fusidic acid resistance rate of S. aureus isolated from Chinese pediatric patients with SSTIs was low, and the genes fusB and fusC were the main resistance determinants. The transposon-like element that contains the fusB gene might participate in the transmission of fusidic acid resistance genes. This is the first report regarding the emergence of fusidic acid-resistant clinical S. aureus isolates in mainland China.

Reduced toxicity with three-dimensional conformal radiotherapy or intensity-modulated radiotherapy compared with conventional two-dimensional radiotherapy for esophageal squamous cell carcinoma: a secondary analysis of data from four prospective clinical trials.

We conducted a retrospective analysis to assess the toxicity and long-term survival of esophageal squamous cell carcinoma patients treated with three-dimensional conformal radiotherapy (3DCRT) or intensity-modulated radiotherapy (IMRT) versus conventional two-dimensional radiotherapy (2DRT). All data in the present study were based on four prospective clinical trials conducted at our institution from 1996 to 2004 and included 308 esophageal squamous cell carcinoma patients treated with 2DRT or 3DCRT/IMRT. Based on the inclusion and exclusion criteria, 254 patients were included in the analysis. Of these patients, 158 were treated with 2DRT, whereas 96 were treated with 3DCRT/IMRT. The rates of ≥Grade3 acute toxicity of the esophagus and lung were 11.5% versus 28.5% (P = 0.002) and 5.2% versus 10.8% (P = 0.127) in the 3DCRT/IMRT and 2DRT groups, respectively. The incidences of ≥Grade 3 late toxicity of the esophagus and lungs were 3.1% versus 10.7% (P = 0.028) and 3.1% versus 5.7% (P = 0.127) in the 3DCRT/IMRT and 2DRT groups, respectively. The 1-year, 3-year and 5-year estimated overall survival rates were 81%, 38% and 34% in the 3DCRT/IMRT group and 79%, 44% and 31% in the 2DRT group, respectively (P = 0.628). The 1-year, 3-year and 5-year local control rates were 88%, 71% and 66% in the 3DCRT/IMRT group and 84%, 66% and 60% in the 2DRT group, respectively (P = 0.412). Fewer incidences of acute and late toxicities were observed in esophageal squamous cell carcinoma patients treated with 3DCRT/IMRT compared with those treated with 2DRT. No significant survival benefit was observed with the use of 3DCRT/IMRT.