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Objective To establish the method of identifying MRSA with Taqman-fluorescence quantitative PCR basing on mecA/nuc/fem B three gene combined detecting.Methods Taking the coagulase positive MRSA,which isolated from the clinical samples and confirmed by VITEK 2 compact microbial analyzer,as the research obj ect,designed mecA/nuc/fem B specific PCR primers and Taqman fluorescent probe by bio-software PrimerPremier 5 and Designer Beacon 7,FAM,HEX and ROX markers were used to label the fluorescent probe at 5’,and the end of 3’was labeled with BHQ1,detected by fluo-rescence quantitative PCR instrment.Results ①1 g/dl gel electrophoresis results showed that the primer’s specificity of mec A/nuc/fem B were good,and molecular weight of the amplification band consistent with the expected molecular weight and no non-specific amplification band.②Three genes were obtained specific amplification in a single tube single channel and single tube multiple channel detection in PCR,and the three gene amplification effect in a single tube single tube single chan-nel and multichannel PCR similar.Conclusion Successfully established a method of multi channel Taqman-probe fluores-cence quantitative PCR identification of MRSA,mec A/nuc/fem B combined detection can effectively differentiate coagulase negative and positive MRSA,improve the accuracy of identification.
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Objective To investigate the clinical distribution situation and drug resistance change of Klebsiella pneumoniae in the Navy General Hospital during 2011‐2015 in order to provide reference for rational use of antibacterial agents in clinic .Methods The clinically isolated Klebsiella pneumoniae in this hospital during 2011‐2015 were selected and performed the analysis on the de‐tection rate ,department distribution ,specimens source ,resistance of antibacterial drugs and change trend of resistance to carbapen‐em antibacterial drugs .Results The number the detected Klebsiella pneumoniae strains and isolation rate during 2011 -2015 showed an increasing trend year by year ,the specimens sources were mainly from 10 departments of intensive care units(ICU) ,hy‐perbaric oxygen department ,respiratory department ,radiation oncology department ,kidney disease department ,etc .;the submitted specimens were dominated by sputum and urine ,accounting for 59 .7% and 21 .4% of submitted specimens ;the drug resistance of Klebsiella pneumoniae during 2011‐2015 showed the increasing trend year by year .Klebsiella pneumoniae had higher resistance rates to piperacillin ,ampicillin ,ampicillin/sulbactam and cefuroxime and had lower resistance rate to amikacin ,imipenem ,meropen‐em and tobramycin ;the resistance rates to imipenem and meropenem were increased year by year ,and pan‐drug resistant Klebsiella pneumoniae showed a rapidly rising trend .Conclusion The drug resistance of Klebsiella pneumonia is serious ,especially carbapene‐ms‐resistant Klebsiella pneumoniae is significantly increased in the recent years ,therefore its drug resistance monitoring should be strengthened for guiding rational drug use in clinic .
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Objective To investigate the distribution of pathogens in intensive care units (ICU ) and their drug resistance . Methods 668 strains of pathogens isolated from specimens from ICU were collected .VITEK 2 Compact automated microbial iden-tification and susceptibility analyzer was utilized to conduct the antimicrobial susceptibility tests .Kirby-Bauer disk diffusion suscep-tibility test(K-B) was employed to conduct the antimicrobial susceptibility test for Gram-negative bacteria cefoperazone/sulbactam . Results 668 strains of pathogens were derived from sputum [434 (65 .0% )] ,blood[83(12 .0% )] ,urine[88(13 .0% )] ,drainage [14(2 .0% )] ,secretions[14(2 .0% )] and other[35(5 .2% )] .Acinetobacter baumannii was the major detected pathogen in Gram-negative bacteria and the resistance rates were over 50% toward other drug excepting levofloxacin ,sulfamethoxazole and amikacin . Staphylococcus Staphylococcus was the major detected pathogen in Gram-positive bacteria and it showed good sensitivity toward ni-trofurantoin ,quinupristin/dalfopristin ,tigecycline and vancomycin .Candida albicans demonstrated the highest detection rate in fun-gi .Conclusion ICU pathogens have drug resistance in serious condition ,and pathogens and drug resistance monitoring should be strengthened .
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Objective To review the prevalence of nosocomial infection and the change of drug resistance in a comprehensive intensive care unit (ICU) and to provide theoretical basis for clinical prevention and treatment.Methods The strains of bacteria and fungi were isolated from ICU and their drug resistance was retrospectively analyzed from Jan.1st,2010 to Dec.31th,2011.Results The main pathogen of nosocomial infection were Gram-negative bacteria ( 73.3％ ),Gram-positive bacteria ( 17.9％ ) and fungi ( 8.7％ ).In bacterial infection,Gram-negative and G-positive bacteria accounted for 80.3％ and 19.7％ respectively.In Gram-negative bacteria,pseudomonas aeruginosa was the major type (21.7％).In Gram-positive bacteria,staphylococcus aureus (31.4％) was most prominent.Drug resistance of bacteria was severe,while that of fungi was mild.Conclusion Bacteria has severe drug resistance and exhibits multi-drug resistance for commonly used antibiotics.The principle of antibiotics application should be mastered and antibiotics should be chosen according to drug-sensitivity tests.
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OBJECTIVE To investigate bacterial contamination of throat spray in different patient groups before and after using it and explore the satisfactory disinfection methods for decreasing the rate of hospital acquired infection.METHODS The methods were used in accordance with Disinfection Technology Criteria.We compared the germicidal effect of five kinds of methods.RESULTS Bacterial contamination was very serious after using throat spray,many bacteria,even pathogens,as well as the normal flora from the region of nasopharynx had been found there.Bacterial contamination was more serious in ENT department patient than in endoscopy patient after using throat sprays.Burning,alcohol,iodophor and disinfectants containing iodine with alcohol were better than oxidized potential water method on disinfection effect.CONCLUSIONS Much attention should be paid to the contamination of throat spray,selecting the most effective disinfection methods,hospital infections caused by invasive manipulation could be prevented when correct methods are adopted.
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OBJECTIVE To study the infectivity of the Vibrio parahaemolyticus isolated from the brine. METHODS Virulence test group: to divide 35 Kunbai mice into 4 subgroups at random: to inject V. parahaemolyticus into the mice′ abdominal cavity of the test group, Staphylococcus aureus or Escherichia coli into those of the positive control groups and aseptic physiological saline into those of negative control group. Wound infection group:to divide 35 SPF mice into 4 subgroups at random after the legs injured: test subgroup (soaked in artificial brine with bacteria ), two positive control subgroups(with S.aureus or E.coli), negative control subgroup(soaked in aseptic physiological saline). To observe the general condition, blood routine, hemoculture, viscera culture of the mice, after 4 days the mice were sacrificed and examined the viscera with pathological analysis. RESULTS Virulence test group: the hemoculture of one mouse was positive after injected the bacteria into its abdominal cavity for 12 hours, and viscera bacterial culture was positive. Wound infection test group:the ratio of wound infection was 100%,the positive ratio of both the hemoculture and the viscera bacteria culture were 10% after the wound soaked in bacteria solution. There were a great deal of neutrophilic granulocytes infiltration and cellulitis in the striated muscles of wound limbs through pathological examination. The infection of severe degree corresponded with the positive control groups, there was no inflammatory reaction in negative control group. CONCLUSIONS The V. parahaemolyticus isolated from the brine has infectivity and makes the wound of the mice be infected and hematoseptic when the concentration reached 10~6 CFU/ml .
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OBJECTIVE To study the pathogenicity of the Vibrio fluvialis isolated from the coastal seawater.METHODS Virulence experiment group:22 Kunming mice were divided into four subgroups in random:V.fluvialis was injected into abdominal cavity in the test subgroup.And Staphylococcus aureus and Escherichia coli were injected into the positive control subgroups,separately and aseptic physiological saline was injected into the negative control group.Wound infection group:22 SPF mice were divided into four supgroups in random after their legs were injured:the experimental supgroup(soaked in artificial seawater with V.fluvialis);the positive control groups(with S.aureus and E.coli,separately);the negative control group(soaked in aseptic artificial seawater).The general condition,blood routine,blood culture,organ culture and wound secretion culture of the mice were observed.The pathological analysis of the mice was taken after sacrifice on the 3rd day.RESULTS In virulence experiment group,among all the 7 mice′s blood culture of V.fluvialis supgroup,5 mice were found V.fluvialis positive after 12 h injection,and 2 mice kept on positive until 24 h.In wound infection group,pathological examination showed there were a large number of neutrophils distributed over the striated muscle of the injured sites and cellulitis formed.CONCLUSIONS The V.fluvialis isolated from the sea water has pathogenicity,and can cause wound) infection and septicemia when the concentration reached 106 CFU/ml.
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OBJECTIVE To investigate the distribution and drug resistance of pathogenic bacteria in blood culture,and provide a basis for clinical treatment.METHODS The blood samples were poured into the blood culture bottles of Beijing Botai Technique Development Center,and cultured with BacT/Alert 3D automated blood culture system.Isolated bacteria were identified by the VITEK system.Drug sensitivity was tested by the BIOMIC.RESULTS From Jan 2003 to Dec 2007,607 strains of pathogenic bacteria were isolated from 4116 clinic blood specimens,the positive rate was 14.8%,Gram-negative bacteria were the most common pathogens then Gram-positive bacteria and fungi.The sensitivity of Gram-negative bacteria to imipenem/cilastatin(TPM) was the best,the next was FEP;the sensitivity of Gram-positive bacteria to teicoplanin(TCN) was the best,Second was VAN.CONCLUSIONS Drug resistance of isolated pathogenic bacteria in blood cultures is very serious.Monitoring the change of pathogens and trends of drug resistance is very important in guiding the clinical use of drug.
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Aim To investigate the effect of total flavonoids of Epimedium sagittatum(TFE)and its metabolites on the proliferation and function expression of newborn rats calvarial osteoblasts(ROB).Methods TFE was supplemented into the culture medium of ROB at 0.2,2,20,100 and 200 mg?L-1 respectively.The serum of rats administered TFES(SRAT) was also added into the medium in a parallel treatment at 2%,4%,8% and 16% respectively.Their effects on cell proliferation and function expression were studied by MTT and the analysis of osteogenic differentiation marks.Results TFE had no effect on cell proliferation and function expression at any concentration.However,2% and 4% SRAT stimulated cell proliferation and,4% SRAT promoted the maturation and function of osteoblast by improving the alkaline phosphatase(ALP) activity,bone gla protein(BGP) secretion,calcium deposition and the number of mineralized nodular structures.Conclusions The effective substance of herba epimedii treating on osteoporosis is not TFE itself but certain of metabolites derived from TFE and the products induced by these metabolites in serum.The metabolites in serum of TFE may enhance the cell function expression and proliferation.