Mei Kang

Resistance trends among clinical isolates in China reported from CHINET surveillance of bacterial resistance, 2005-2014

With the aim of gathering temporal trends on bacterial epidemiology and resistance from multiple laboratories in China, the CHINET surveillance system was organized in 2005. Antimicrobial susceptibility testing was carried out according to a unified protocol using the Kirby-Bauer method or automated systems. Results were analyzed according to Clinical and Laboratory Standards Institute (CLSI) 2014 definitions. Between 2005 and 2014, the number of bacterial isolates ranged between 22,774 and 84,572 annually. Rates of extended-spectrum β-lactamase production among Escherichia coli isolates were stable, between 51.7 and 55.8%. Resistance of E. coli and Klebsiella pneumoniae to amikacin, ciprofloxacin, piperacillin/tazobactam and cefoperazone/sulbactam decreased with time. Carbapenem resistance among K. pneumoniae isolates increased from 2.4 to 13.4%. Resistance of Pseudomonas aeruginosa strains against all of antimicrobial agents tested including imipenem and meropenem decreased with time. On the contrary, resistance of Acinetobacter baumannii strains to carbapenems increased from 31 to 66.7%. A marked decrease of methicillin resistance from 69% in 2005 to 44.6% in 2014 was observed for Staphylococcus aureus. Carbapenem resistance rates in K. pneumoniae and A. baumannii in China are high. Our results indicate the importance of bacterial surveillance studies.

Microbiologic study of the pathogens isolated from wound culture among Wenchuan earthquake survivors

On May 12, an unprecedented earthquake struck Wenchuan County, Sichuan, China, and 1823 victims were admitted to West China Hospital, Sichuan, China. A total of 464 nonduplicate clinical isolates from wounds of earthquake victims were analyzed. The results show that the most common pathogen was Staphylococcus aureus, but only 24.4% of the total isolates were Gram-positive bacteria, and 73.2% were Gram-negative bacteria. The isolates were significantly different from isolation pattern of 2004 tsunami in Thailand. The isolation rates of extended-spectrum beta-lactamase producer and pandrug-resistant Acinetobacter baumannii in this study might increase the risk of nosocomial infection. In this situation, clinical microbiologists, infection control staff, and administration decision makers should pay high attention to prevent disaster-associated nosocomial infections.

Spread of imipenem-resistant Acinetobacter baumannii of European clone II in Western China

The aim of this study was to investigate the distribution of resistance genes and the clonal relationship amongst imipenem-resistant Acinetobacter baumannii isolates from ten hospitals in Western China as well as to compare the molecular epidemiological data with those of isolates from two hospitals in Hangzhou and Beijing. Genes encoding OXA carbapenemases, metallo-β-lactamases, AmpC cephalosporinase and carbapenem resistance-associated outer membrane protein (CarO) were screened using polymerase chain reaction (PCR) and sequencing. PCR mapping was performed to determine whether insertion sequence ISAba1 elements preceded OXA carbapenemases and AmpC cephalosporinase. International clonal lineages were identified by sequence type multiplex PCR. Multilocus sequence typing (MLST) was performed to determine the sequence types (STs), and then eBURST algorithm was applied to assign clonal complexes (CCs). In this study, dissemination of acquired ISAba1 preceding the bla(OXA-23-like) gene was the predominant enzymatic resistance mechanism amongst 272 imipenem-resistant isolates. Five isolates harboured the carO gene disrupted by insertion of ISAba1 and three isolates lacked carO. All of the 36 representative isolates belonged to European clone II. Ten STs, including three novel types, were identified. These STs were clustered into CC92 and two distinct singletons. These observations suggest that imipenem-resistant A. baumannii of European clone II, which carries acquired ISAba1 preceding the bla(OXA-23-like) gene and belongs to CC92, has spread within Western China.

Increasing imipenem resistance and dissemination of the ISAba1-associated blaOXA-23 gene among Acinetobacter baumannii isolates in an intensive care unit

The antibiotic susceptibility of Acinetobacter calcoaceticus-Acinetobacter baumannii complex strains recovered from the intensive care unit (ICU) of West China Hospital, Sichuan, PR China, from 2006 to 2009 was investigated. The identification of A. baumannii and analysis of carbapenemase-encoding genes and their relationship with ISAba1 were performed by PCR. Furthermore, a DiversiLab repetitive extragenic palindromic sequence-based PCR (rep-PCR) microbial typing system and a multilocus sequence typing (MLST) scheme were applied to assess the genetic relationship of the isolates. The results showed that the antibiotic susceptibility of the A. calcoaceticus-A. baumannii complex isolates changed and imipenem resistance increased rapidly between 2006 and 2009. The blaOXA-51-like and ISAba1-associated blaOXA-23 genes were prevalent in the imipenem-resistant A. baumannii isolates. However, the blaOXA-58-like gene was found in only one isolate and no metallo-β-lactamase genes were detected. The representative multidrug-resistant A. baumannii isolates were identified as one cluster by rep-PCR fingerprinting and belonged to the clonal complex 92 (CC92) according to MLST. These findings indicate a situation of increasing resistance and wide distribution of class D β-lactamase genes, especially the acquired ISAba1-associated blaOXA-23 gene, in A. baumannii isolates in the ICU of West China Hospital, probably caused by expansion of the CC92 clone.

Molecular characterisation of clinical Cryptococcus neoformans and Cryptococcus gattii isolates from Sichuan province, China

Previous reports on the molecular characteristics of clinical isolates of Cryptococcus species in China have focused on isolates from southeast China. To obtain a more detailed molecular epidemiology, a total of 92 cryptococcal isolates were collected from Sichuan province. A total of 24 isolates from 12 other provinces were collected for comparative study. Genotypes and mating types of 116 Cryptococcus isolates were determined. Among the 116 isolates, 43 isolates (19 isolates from Sichuan and 24 isolates outside of Sichuan) were analysed by multi‐locus sequence typing (MLST). All 116 clinical isolates were mating type α. Most isolates (114/116) were molecular type VNI and the remaining two isolates were VGI and VGII respectively. MLST results revealed five sequence types (STs) of C. neoformans including two novel STs, with most isolates identified as ST5. The two C. gattii isolates identified in our study were ST44 and ST159. Based on our report and previous studies, there are 15 C. neoformans STs in China which can be divided into three subgroups. The C. gattii isolate from Sichuan could be a scattered subtype of VGII (ST44). Our findings demonstrated that C. neoformans isolates in Sichuan are genetically homogeneous, and ST5 is the epidemic clone of C. neoformans in China.

Enhancing pili assembly and biofilm formation in Acinetobacter baumannii ATCC19606 using non-native acyl-homoserine lactones

BackgroundQuorum Sensing (QS) systems influence biofilm formation, an important virulence factor related to the bacterial survival and antibiotic resistance. In Acinetobacter baumannii, biofilm formation depends on pili biosynthesis, structures assembled via the csuA/BABCDE chaperone-usher secretion system. QS signaling molecules are hypothesized to affect pili formation; however, the mechanism behind this remains unclear. This study aimed to demonstrate the possible role of QS signaling molecules in regulating pili formation and mediating the ability to form biofilms on abiotic surfaces.ResultsReal-time quantitative PCR analysis showed the expression of the csuA/BABCDE genes distinctly increased when co-cultured with C6-HSL (P < 0.05). Under the same experimental conditions, expression of BfmS and BfmR was significantly higher than the control strain (P < 0.05). A subsurface twitching assay showed a switch from a small to a large and structured clone that may result from enhanced twitching motility (P < 0.05). Transmission electron microscopy analysis of cells lifted from a MH broth co-cultured with C6-HSL showed more abundant pili-like structures than the control strain. We then tested the idea that the addition of a QS signal, and therefore induction of chaperone-usher secretion system genes, provides a greater benefit at higher biofilm densities. An assay for the total fluorescence intensity of the biofilm using Confocal Laser Scanning Microscopy revealed an obvious increase.ConclusionOur study demonstrated that, increased transcription of the BfmS and BfmR genes, QS signaling molecules enhance the expression of the chaperone-usher secretion system, and this expression is required for twitching motility in A. baumannii. The concomitant pili expression and strain twitching allowed A. baumannii to attach easily to abiotic surfaces and form biofilms at an earlier timepoint.

Antimicrobial susceptibility of clinical isolates from earthquake victims in Wenchuan

On 12 May 2008, an earthquake measuring 8.0 on the Richter scale struck Wenchuan County, Sichuan, China. Between 12 May and 11 June, 1823 victims were hospitalized in West China Hospital. These patients were severely injured, and most of their wounds were contaminated. Here, the results of bacteriological identification and antibiotic susceptibility testing of 725 non-duplicate isolates from earthquake victims are presented. Gram-negative bacilli were most frequently isolated (71.3%). Only 18.9% of isolates were Gram-positive bacteria; Candida spp. accounted for 9.7%, and Gram-negative cocci for 0.1%. After anaerobic culture, four Clostridium sordellii strains and one Clostridium bifermentans strain were isolated from deep wounds. Specimen culture from earthquake victims revealed a spectrum of pathogens and antibiotic susceptibilities that was different from that usually encountered in West China Hospital, especially concerning methicillin-resistant Staphylococcus aureus, extended-spectrum beta-lactamase producers, and multidrug-resistant (MDR) non-fermenting Gram-negative bacilli. The pathophysiology of the injuries in earthquake victims was different from that in the patients who were not earthquake victims. A combination of environmental bacteria with a high proportion of Gram-negative bacteria was often observed in the earthquake victims. Approximately 26% of all earthquake victims were shown to be carriers of MDR microorganisms. Therefore, appropriate microbiological assessment upon admission, and identification of patients to be put in quarantine, is of paramount importance.

A mixed infection of Leuconostoc lactis and vancomycin-resistant Enterococcus in a liver transplant recipient.

Bacterial infection in patients who have undergone liver transplantation is a major complication of the procedure. Leuconostoc spp. are important pathogenic bacteria in individuals with poor immune function, especially transplant patients. In this report, we describe the case of a 45-year-old Asian male liver transplant recipient who was initially preliminarily diagnosed with infection with Leuconostoc pseudomesenteroides by using the microbial tests of the VITEK 2 system and the aesculin hydrolysis test, and with vancomycin-resistant Enterococcus. Subsequently, the Leuconostoc isolate was identified as Leuconostoc lactis by 16S rRNA gene partial sequencing. In this paper, we discuss our identification of L. lactis based on physiological characteristics and molecular methodology. Accurate identification of these infections is important for the outcome; use of 16S rRNA gene sequence analysis offers a rapid and precise diagnostic approach. Administration of the drug linezolid may be useful for the treatment of both Leuconostoc spp. and vancomycin-resistant Enterococcus infections. We suggest that clinical analysts should use molecular methods in addition to biochemical tests in order to identify Leuconostoc at the species level more accurately.

Molecular epidemiological analysis of methicillin-resistant Staphylococcus aureus isolates from a medical intensive care unit: a comparison of nasal and clinical isolates.

Background:The control of nosocomial transmission of methicillin-resistant Staphylococcus aureus (MRSA) represents a significant challenge to infection control professionals. Nasal carriage colonization by MRSA plays a crucial role in the epidemiology and pathogenesis of this infection. Methods:Patients in the medical intensive care unit (ICU) between November 2010 and March 2011 were swabbed when hospitalized, reswabbed 1 week later and for a third time, when they were discharged from the ICU. All swabs were examined within 2 hours of collection using ChromID MRSA-Select agar plates to detect MRSA. Positive specimens were determined to have the mecA and femB gene through amplification with duplex polymerase chain reaction. Repetitive element sequence–based polymerase chain reaction was used to investigate the epidemiological types of MRSA isolates in the third screening and clinical isolates obtained from 2007 to 2010 in West China Hospital. A comparison of molecular types was performed to investigate the genetic relationship between nasal and clinical isolates. Results:After the third screening, 16 nasal MRSA isolates were identified. Epidemiological analysis revealed that 16 nasal MRSA isolates and 37 clinical MRSA isolates differentiated into 2 clusters, comprising 9 subclusters. Of the 16 nasal strains, 11 (68.8%) belonged to subcluster I of cluster I; 3 of 9 subclusters consisted of both nasal and clinical isolates, while 4 of 9 subclusters consisted of clinical isolates and only 2 of 9 consisted of nasal isolates. Conclusions:Our study indicated a high degree of genetic relatedness between nasal and clinical MRSA isolates. The molecular typing of MRSA is critical for controlling the nosocomial transmission of this pathogen in ICU setting and defining a nosocomial infection control policy.

The microbiological characteristics of patients with crush syndrome after the Wenchuan earthquake

Abstract To improve the treatment of infectious diseases in patients with crush syndrome, we analyzed the results of bacteriological examinations, including antimicrobial susceptibility patterns, of samples taken from patients with crush syndrome admitted to West China Hospital, Sichuan University, after the Wenchuan earthquake. A total of 210 non-replicate clinical isolates were recovered from 42 of the 66 earthquake victims with crush syndrome. Their mean age was 26.9 ± 15.7 y and 40 of them were male. The length of hospital stay was 14.0 days. Wound, blood, sputa, urine and catheter sample specimens were examined. Gram-negative bacilli, Gram-positive bacteria and fungi accounted for 72.4%, 20.0% and 7.6% of the isolates, respectively. Acinetobacter baumannii and Pseudomonas aeruginosa were the major isolates from wounds. Of the isolated strains, 92.8% occurred at >48 h following admission to the hospital, and most of these agents were common isolates in our hospital. Furthermore, 40.9% of these patients were carriers of multidrug-resistant (MDR) microorganisms. Therefore, we can conclude that patients with crush syndrome may have an increased risk of hospital-acquired infection (HAI), suggesting that it is important to select effective antibiotics to control HAI in a timely manner according to the microbiological data.