1.Correct understanding of superbugs with New Delhi metallo-beta-lactamase-1
Chinese Journal of Laboratory Medicine 2010;33(12):1109-1111
The term "superbugs" refers to microbes with resistance to almost all antibiotics specifically recommended for the treatment. The superbugs disseminating worldwide included methicillinresistant Staphylococcus aureus ; vancomycin-resistant Staphylococcus aureus ; vancomycin-resistant Enterococcus ;carbapenem-resistant Gram-negative bacillus. Due to abuse of antibiotics, novel "superbug"emerged gradually, which demands optimizing antibiotic use, reinforcing surveillance of "superbugs" as well as nosocomial infection control.
2.Joint efforts are required from the clinic and laboratory to improve the level of antimicrobial therapy
Chinese Journal of Laboratory Medicine 2012;35(8):679-681
The key to improve the clinic antimicrobial therapy results is correct diagnosis of infection,pathogens and antibiotic resistance.To deal with increasingly complex pathogens and drug resistance,the clinic and lab should cooperate and communicate as often as possible,improve the level of pathogen diagnosis,interpretate of the microbiological examination report correctly and explore the mechanisms and treatment of antimicrobial resistant pathogens.Besides,they should make full use of the inflammatory markers to further enhance anti-infective levels.
3.To promote studies on community-associated methicillin-resistant Staphylococcus aureus in China
Chinese Journal of Clinical Infectious Diseases 2016;9(4):294-298
Community-acquired methicillin-resistant Staphylococcus aureus ( CA-MRSA ) has been increasingly frequently isolated from patients worldwide, and is an emerging threat to public health.CA-MRSA strains differ from hospital-acquired MRSA strains in their epidemiologies, clinical features and genetic backgrounds, and the predominant CA-MRSA strains vary between geographic settings.This paper reviews literatures on the definition, epidemiology, clinical manifestations, treatment and virulent factors of CA-MRSA or CA-MRSA infection, and points out that we should further promote studies on epidemiology, molecular genetic properties of CA-MRSA and clinical management of CA-MRSA infection.
4.Cloning and sequencing of the gene encoding extended-spectrum ?-lactamases SHV-12
Weilin ZHOU ; Yagang CHEN ; Yunsong YU
Chinese Journal of Infectious Diseases 2001;0(05):-
Objective To analyse the encoding gene sequence of extended spectrum ? lactamase produced by Klebsiella pneumoniae E95 strain in Zhejiang Province and identify its ESBLs subtype. Methods The gene of ESBLs produced by E95 strain was amplified by PCR. The purified PCR product was cloned into pGEM Teasy vector and then sequenced by Sanger's dideoxy chain termination composition method. Results The encoded gene of ESBLs was identified as SHV by PCR. Its PCR product had 812 nucleotides. It had the same gene sequence as the gene encoding SHV 12 discovered in Swiss. Conclusions The ESBLs produced by Klebsiella pneumoniae E95 strain isolated from a patient in Zhejiang Province is SHV 12.
5.Establishment of mice pneumonia model with imipenem-resistant Acinetobacter baumannii
Daojun YU ; Yunsong YU ; Xiang FANG ; Xianjun WANG ; Yijian PAN
Chinese Journal of Laboratory Medicine 2010;33(8):771-775
Objective To construct the mice pneumonia model with imipenem-resistant Acinetobacter baumannii and provide experimental model in anti-pan-resistant Acinetobacter baumannii therapy study. Methods A total number of 120 4-week-old BALB/C male mice were randomly selected and divided into three groups including micro-intratracheal injection, ultrasonic atomizing and nasal dripping. The mice were treated with methotrexate to induce hypo-immunity in every group. These BALB/C mice of normal immunity and hypo-immunity were infected through Imipenem-resistant Acinetobacter baumannii by microintratracheal injection, ultrasonic atomizing and nasal dripping, respectively. The morbidity, mortality,bacterial clearance rate and histopathology in lung were determined. Results The morbidities of BALB/C mice with hypo-immunity infected by micro-intratracheal injection and ultrasonic atomizing achieved 100%(30/30), while the mortalities were 100% (10/10) and 33.3% (3/10), respectively. Mice in two groups above displayed the influx of neutrophils, lymphocytes and macrophages in the peri-bronchial and alveolar interstitial space 12-24 h after pulmonary infection. In addtion, the mice in micro-intratracheal injection group displayed coUapse of partial alveolar walls, formation of abscesses and bacterial colonies in alveoli. While the lung pathology in mice of ultrasonic atomizing group was characterized by cell degeneration in some regions in the lungs, slight relaxation, congestion in alveolar wall vessels and normal of bronchial and alveolar tissue 24 h after inoculation. Degeneration in peri-tracheal and peri-bronchial areas was observed 24-48 h after inoculation, along with highly expanded pulmonary blood vessels and edems. The inflammation was reduced at 48 hours. There was no obvious pulmonary infection in BALB/C mice with hypo-immunity by nasal dripping with mortality of 0% (0/10) and no significant histopathologic change in lungs. Conclusions BALB/C mice with hypo-immunity pneumonia model with Imipenem-resistant Acinetobacter baumannii can be conducted by micro-intratracheal injection or ultrasonic atomizing, but the latter has the advantages of high-productivity, easy-operation, low-cost, time-saving and usefulness. Mice with normal immunity are not susceptible to imipenem-resistant Acinetobacter baumannii.
6.Study on genic expression of ampC in Serratia marcescens
Daojun YU ; Tianmei ZHOU ; Xiaoqin DONG ; Yunsong YU ; Yagang CHEN
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objectives To investigate the properties and distributions of ampC gene among different drug-resistant strains of Serratia marcescens,and the relationship of control gene ampR with AmpC enzymes′ expressions.Methods According to the results of inducting experiment with 1/2 MIC of beta-lactam antibiotics (CTX),three-dimensional testing and isoelectric focusing electrophoresis testing,143 strains of S.marcescens were classified into three groups:including induction group, continuous low-production group and hyperproduction group. In each group, the sequences of ampC and ampR genes were amplified using the method of PCR. The products of PCR were analyzed. The plasmid-mediated beta-lactamases were detected using the method of conjugation experiment.Results Among 143 strains of S.marcescens, the continuous low -production strains, induction strains and hyperproduction strains were 14,103,and 18, respectively.125 and 99 strains were ampC and ampR gene positive, respectively.The detection rate of ampR in hyperproduction group was lower than other groups.5 sites of ampC genes and 4 sites in the Open Reading Frame (ORF) of ampR gene were easily mutated in 5 induction strains and 2 hyperproduction strains.Conclusions The production of inducing drug-resistance of some S.marcescens might be related to mutation of ampC gene encoding AmpC beta-lactamases and the ORF mutation in ampR. The continuous hyperproduction drug-resistance had something to do with deletion mutation in ampR in segmental hyperproduction strains.The plasmid-mediated AmpC enzymes hadn′t been found in S.marcescens.
7.Investigation on the genotyping of Acinetobacter baumannii by DiversiLab system
Yanyan QU ; Wenfei WANG ; Hua ZHOU ; Yiqi FU ; Yunsong YU
Chinese Journal of Laboratory Medicine 2010;33(5):425-429
Objective To evaluate the usefulness of DiversiLab system for genotyping of Acinetobacter baumannii.Methods Fifty-eight non-duplicated clinical Acinetobacter baumannii isolated from 15 cities in China in 2005 were typed by rep-PCR-based DiversiLab system.The results were compared with those of PFGE and multilocus sequence typing. Simpson's index of diversity was used to compare the discriminatory power among DiversiLab system, PFGE and MLST.Results Fifty-eight Acinetobacter baumannii isolates were differentiated into 5 clusters and 25 unique types by DiversiLab system. MLST identified 35 distinct sequence types, which fell into one clone complex of CC22 and 35 singletons, while PFGE resolved 5 pulsotypes and 34 unique types.Simpson's diversity indices for DiversiLab system, MLST and PFGE were O.876, O.944 and 0.961, respectively.Conclusions The discriminatory power of DiversiLab system is lower than that of PFGE and MLST.But as a simple, fast and reproducible typing method, it could be used as a first-line typing tool for the analysis of a large number of isolates.
8.The distribution of 16S rRNA methylase genes in KPC-producing Klebsiella pneumoniae strains
Liying LU ; Weili ZHANG ; Qing YANG ; Hua ZHOU ; Yunsong YU
Chinese Journal of Clinical Infectious Diseases 2009;2(2):71-73
Objective To investigate the distribution of 16S rRNA methylase genes in Klebsiella pneumoniae strains producing Klebsiella pneumoniae ealbapenenase type 2(KPC-2).Methods A total of 37 Klebsiella pneumoniae isolates producing KPC-2 were collected.The minimal inhibitory concentrations (MICs)of these strains to amikacin,gentamyein and netilmicin were determinated by agal dilution method.Six 16S rRNA methylase genes(armA,rmtA,rmtB,rmtC,rmtD and npmA)were detected by PCR.Results The resistant rates to amikacin,gentamycin and netilmicin were 97.3%(MIC50≥1024μg/mL).Among those resistant strains,8 were armr/A positive,25 were rmtB positive,4 were both armA and rmtB positive.and no other 16S rRNA methylase genes were found.The total positive rate of 16S rRNA methylase genes was 78.4%(29/37).Conclusion 16S rRNA methylase genes armA and rmtB ale prevalent in Klebsiella pneumoniae strains producing KPC-2.
9.Clinical analysis of Stenotrophomonas maltophilia septicemia in 15 patients with liver transplantation
Suqiu HU ; Qing YANG ; Xiaoxing DU ; Yunsong YU ; Lanjuan LI
Chinese Journal of Clinical Infectious Diseases 2008;1(4):210-213
Objective To investigate the related clinical factors and homology of strains in Stenotrophomonas maltophilia (S. Maltophilia) infections in 15 patients with liver transplantation. Methods Fifteen patients with S. Maltophilia infection from September to December 2006 were enrolled and their clinical data were collected. Minimal inhibitory concentrations (MICs) of 10 antimierobial agents against S. Maltophilia were determined by Etest strips. Antibiogram was carried out by resistance analysis assembly with WHONET 5 software. The genomic DNA of all the isolates was digested with Xbal and subjected to pulse-field gel electrophoresis (PFGE). Results All patients received mechanical ventilation during the treatment and had a history of long-term use of extended-spectrum β-lactamases and quinolones. MICs of 10 antimicrobial agents indicated that S. Maltophilia were susceptible to several antimicrobial agents including compound sulfamethoxazole and ciprofloxacin, but the best active agent against these resistant isolates was minocycline in vitro. The results of all 15 S. Maltophilia antibiograms were accordance with PFGE patterns. All 15 S. Maltophilia isolates were classified as 2 PFGE patterns: 9 for pattern A and 6 for pattern B. Conclusion Mechanical ventilation might be associated with the S. Maltophilia septicemia in patients with liver transplantation.
10.Structures of resistance transposons and multilocus sequencing typing in vancomycin-resistant enterococci strains isolated from five hospitals in Hangzhou
Tingting QU ; Yunsong YU ; Zeqing WEI ; Yagang CHEN ; Lanjuan LI
Chinese Journal of Laboratory Medicine 2008;31(6):628-634
Objective To determine the structures of resistance transposons and muhilocus sequencing typing(MLST)in the vancomycin resistant enterococcus(VRE).Methods Twenty-one VRE strains were isolated from five hospitals in Hangzhou.The resistance to antimicrobial agents was determined by Etest.Polymerase chain reaction(PCR),conjugation,plasmid extract,transposon structures,pulse field gel electrophoresis(PFGE),muhilocus sequencing typing(MLST),and multiple-locus variable-number tandem repeat analysis(MLVA)were carried out.Results All of the 21 VRE strains harbored the vanA gene.These strains were divided into 10 PFGE types,7 sequence types(STs)and 5 MLVA types.All of these VRE strains were susceptible to linezolid and tigecycline.The vanA genes in two VRE strains were located in transposon Tnl546,and those in the other 19 VRE strains were located in transpeson Tnl546- like,with ISl485 inserted in vanXY.Vancomycin resistance of 1 8 VRE isolates was transferred by filter mating. All of these conjugants had a plasmid containing a molecular size of about 54 000 bo.Conclusions These 21 VRE strains were all caused by the vanA gene and divided into 7 MIST types.A novel trasnposon was detected.Most of these VRE isolates belonged to the clonal complex(CC17)by MIST,which was the hospital-adapted and pandemic VRE clonal complex.