Dingqiang Chen

Resistance integrons: class 1, 2 and 3 integrons

As recently indiscriminate abuse of existing antibiotics in both clinical and veterinary treatment leads to proliferation of antibiotic resistance in microbes and poses a dilemma for the future treatment of such bacterial infection, antimicrobial resistance has been considered to be one of the currently leading concerns in global public health, and reported to widely spread and extended to a large variety of microorganisms. In China, as one of the currently worst areas for antibiotics abuse, the annual prescription of antibiotics, including both clinical and veterinary treatment, has approaching 140 gram per person and been roughly estimated to be 10 times higher than that in the United Kingdom, which is considered to be a potential area for the emergence of “Super Bugs”. Based on the integrons surveillance in Guangzhou, China in the past decade, this review thus aimed at summarizing the role of integrons in the perspective of both clinical setting and environment, with the focus on the occurrence and prevalence of class 1, 2 and 3 integrons.

Antimicrobial resistance investigation on staphylococcus strains in a local Hospital in Guangzhou, China, 2001-2010

A retrospective study was conducted on 1,739 Staphylococcus isolates from the First Affiliated Hospital of Jinan University (FAHJU) in Guangzhou during 2001-2010. With the exception of teicoplanin and vancomycin, antimicrobial resistance was commonly observed among the isolates examined, with high resistance rates for β-lactamases (94.0% and 73.7% for penicillin and oxacillin) and resistance percentages for cefoxitin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, trimethoprim-sulfamethoxazole, and tetracycline ranging from 83.9% to 19.4%. Two hundred sixty-three of the 1,739 isolates were subjected to SCCmec typing and 42 to MLST, spaA, and coa typing. ST239-MRSA-III was prevalently identified along with one distinct coa type HIJKL and 2 spaA types (WGKAOMQ-t037 and WGKAQQ-t030). Class 1 integrons were commonly detected (31.6%), although none of the integron-positive MRSA strains had been isolated since 2009. The widespread detection of integron-based antimicrobial resistance determinants may further contribute to the emergence of superbugs.

Chromogenic media for MRSA diagnostics.

For the past decade, a number of chromogenic media for methicillin-resistance Staphylococcus aureus (MRSA) detection have been developed and applied, including Oxoid Brilliance™ MRSA, CHROMagar™ MRSA, BBL™ CHROMagar™ MRSA, MRSASelect and chromID MRSA. The advantages of these chromogenic media offers direct detection of visible staphylococcal colonies, coupled with the use of chromogenic enzymatic substrates that can be hydrolyzed by S. aureus to confirm species or strain identification. BBL™ CHROMagar™ MRSA and MRSASelect are designed for detection of nasal colonization by MRSA, while CHROMagar™ MRSA, Oxoid Brilliance™ MRSA and chromID MRSA are readily applied in bacterial screening. This review summarizes the characteristics, principles and capacities of these selective media, and focuses on comparison of different chromogenic media.

Current methodologies on genotyping for nosocomial pathogen methicillin-resistant Staphylococcus aureus (MRSA)

Methicillin-resistant Staphylococcus aureus (MRSA) is a common pathogen in hospitals and the community. As the rapid spread and wide distribution of antimicrobial resistance (such as MRSA), treatment for infectious diseases caused by microorganisms has become a vital threat. Thus, early identification and genotyping are essential for further therapeutic treatment and the control of rapid expansion of MRSA. In combination with applications and data feedbacks, this review focused on the currently available molecular-based assays on their utility and performance for rapid typing of MRSA, especially on effective molecular-based methods. Besides, a common mobile element SCCmec and prevalence of HA-MRSA, LA-MRSA and CA-MRSA were introduced in this review in order to provide a more complete profile of MRSA.

Evaluation and application of molecular genotyping on nosocomial pathogen-methicillin-resistant Staphylococcus aureus isolates in Guangzhou representative of Southern China

Staphylococcus aureus is recognized as a typical food-born pathogen in food safety, and its rapid detection and typing methods are in desperate need. To study their evolution characteristics and drug resistance condition, Staphylococci isolates were subject to the investigation. Strain identification were subjected to standard procedures (colony morphology, Gram staining, Vitek 2 automated system and the API commercial kit), and fingerprinting was then performed with RAPD-PCR amplification. In this study, 179 isolated staphylococci were identified as MRSA. In addition, Staphylococci isolates were subjected to three different fingerprinting system (ERIC2, AP1 and AP7) and then genotyped based on significant diversities. Nosocomial epidemiology was then conducted according to the fingerprint result. To sum up, the RAPD method subjected for MRSA rapid genotyping have a broad application prospect in food safety and epidemiology.

Complete sequence of pBM413, a novel multidrug resistance megaplasmid carrying qnrVC6 and blaIMP-45 from pseudomonas aeruginosa

This study aimed to characterise a novel multidrug resistance megaplasmid carrying qnrVC6 and blaIMP-45 from Pseudomonas aeruginosa strain Guangzhou-Pae617 isolated from a patient hospitalised in Guangzhou, China, in 2012. The plasmid pBM413 has a length of 423 017 bp and an average G + C content of 56.41%. A qnrVC6 gene flanked by two copies of insertion sequence (IS) elements ISCR1, a multiresistance class 1 integron In786 containing aacA4-blaIMP-45-blaOXA-1-catB3 cassettes, an armA gene, and an aphA7 gene flanked by two copies of IS26 were identified. To our knowledge, this is the first identification of a qnrVC6 gene in P. aeruginosa.

Pathogenic features and characteristics of food borne pathogens biofilm: Biomass, viability and matrix

Biofilm is a ubiquitous growth pattern of bacterial species survival but is notorious for its threat on public health and food contamination. Extensive studies of the biofilm structure, formation, quantification, quorum sensing system and underlying control strategies have been reported during the past decades. Insightful elucidation of the pathogenic features and characteristic of bacterial biofilm can facilitate in devising appropriate control strategies for biofilm eradication. Therefore, this review mainly summarized the pathogenic features of biofilms from food borne microorganisms, including the biomass (which could be quantified using crystal violet and fluorogenic dye Syto9 assays), viability (which could be determined by tetrazolium salts, fluorescein diacetate, resazurin staining and alamar blue assays) and matrix (which are commonly detected by dimethyl methylene blue and wheat germ agglutinin assays). In addition, three features were further compared with its particular benefits in specific application.

Effect of aminoglycosides on the pathogenic characteristics of microbiology

Infections caused by pathogen remain to be one of the most important global health issues, and scientists are devoting themselves to seeking effective treatments. Aminoglycoside antibiotics are one kind of widely used antibiotics because of the good efficiency and broad antimicrobial-spectrum. However, it causes some unexpected effects on the pathogenic characteristics of microbiology during the treatment, such as drug resistance and biofilm promotion. Drug resistance is partly due to antibiotics abuse. Simultaneously, aminoglycoside is documented to make divergent effects on biofilm based on their concentrations. Here, we review the mechanism of drug resistance caused by long-term use of aminoglycoside antibiotics, the effects of antibiotic concentration on biofilm formation and the negative effects on intestinal flora to provide theoretical supports for rational use of antibiotics.

The novel loop-mediated isothermal amplification based confirmation methodology on the bacteria in Viable but Non-Culturable (VBNC) state

As a self-protection mechanism, the viable but non-culturable (VBNC) state provides the ability against conventional detection methods among various foodborne pathogens. The ability of forming colonies is lost while metabolism is still maintaining in VBNC state cells. Recently, ethidium monoazide (EMA) and propidium monoazide (PMA) have been widely applied on the detection of foodborne pathogens in VBNC state. Combined with loop-mediated isothermal amplification (LAMP), the PMA/EMA-LAMP showed a significant priority in high sensitivity, specificity and rapidity over conventional PCR based assays. Particularly, PMA/EMA-LAMP has been proved as an effective method in the detection of Escherichia coli, Vibrio parahaemolyticus and Staphylococcus in VBNC state. Based on the current investigations, the VBNC mechanism and current detection method for VBNC-state foodborne pathogens were introduced and discussed in this review.

Loop-mediated isothermal amplification assays for screening of bacterial integrons

BackgroundThe occurrence and prevalence of integrons in clinical microorganisms and their role played in antimicrobial resistance have been well studied recently. As screening and detection of integrons are concerned, current diagnostic methodologies are restricted by significant drawbacks and novel methods are required for integrons detection.ResultsIn this study, three loop-mediated isothermal amplification (LAMP) assays targeting on class 1, 2 and 3 integrons were implemented and evaluated. Optimization of these detection assays were performed, including studing on the reaction temperature, volume, time, sensitivity and specificity (both primers and targets). Application of the established LAMP assays were further verified on a total of 1082 isolates (previously identified to be 397 integron-positive and 685 integron-negative strains). According to the results, the indispensability of each primer had been confirmed and the optimal reaction temperature, volume and time were found to be 65°C, 45 min and 25 μL, respectively. As application was concerned, 361, 28 and 8 isolates carrying intI1, intI2 and intI3 yielded positive amplicons, respectively. Other 685 integron-negative bacteria were negative for the integron-screening LAMP assays, totaling the detection rate and specificity to be 100%.ConclusionsThe intI1-, intI2- and intI3-LAMP assays established in this study were demonstrated to be the valid and rapid detection methodologies for the screening of bacterial integrons.