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Objective To establish a polymerase spiral reaction (PSR) method for rapid detection of influenza A/H1N1 virus.Methods Six sets of primers were designed for influenza A/H1N1 virus HA gene, and the results were determined with real time kinetic turbidimetric assay and colorimetry method.Results and Conclusion The best primers were selected from six sets of primers, and the best temperature was determined as 65 degrees Celsius.Further experiments showed that the best primer had good specificity for detection of influenza A/H1N1 virus,without cross reactions with 14 other respiratory tract pathogenic nucleic acids.The sensitivity was up to 100 copies,and consistent with that of PCR.So a PSR method is established for rapid detection of the influenza A/H1N1 virus, which is simple, quick, highly specific and sensitive,and especially applicable to field and grass-roots units.
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Being an important clinical fungal pathogen,Aspergillus (A.)fumigatus can cause fatal invasive fungal infections.Azoles are the first line drugs in treating various Aspergillus-caused diseases.Worldwidely,reports related to azole resistance in A.fumigatus have been increasing which posing a threat on the effectiveness of clinically used azole and agricultural fungicides.Currently,it has become an important public health issue.In this review,we summarize findings from literature regarding the following areas:the occurrence of azole resistance in A.fumigatus,the molecular mechanisms of resistance,contributing factors for the emergence of azole resistance,evolution of resistant strains and related control and prevention measures.
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Being an important clinical fungal pathogen,Aspergillus (A.)fumigatus can cause fatal invasive fungal infections.Azoles are the first line drugs in treating various Aspergillus-caused diseases.Worldwidely,reports related to azole resistance in A.fumigatus have been increasing which posing a threat on the effectiveness of clinically used azole and agricultural fungicides.Currently,it has become an important public health issue.In this review,we summarize findings from literature regarding the following areas:the occurrence of azole resistance in A.fumigatus,the molecular mechanisms of resistance,contributing factors for the emergence of azole resistance,evolution of resistant strains and related control and prevention measures.
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Objective To establish a multiplex loop-mediated isothermal amplification(mLAMP)method for simultaneous detection of Salmonella,Vibrio parahaemolyticus (VPH)and Listeria monocytogenes (LM).Methods Three sets of mLAMP primers were designed to specifically target bcfD of Salmonella and tlh of VPH and iap of LM.The respective single LAMP assay of the three kinds of bacteria was developed,and the ratio of primer concentration was optimized to develop a multiplex LAMP system.The specificity and sensitivity of multiplex LAMP were observed.Results Turbidity monitoring results in real time suggests that the mLAMP was highly specific and amplification could be obtained within 45 min under isothermal conditions.The sensitivity of this mLAMP was found to be 300 fg/μl genomic DNAs for Salmonella and 4.2 pg/μl for VPH and 4.5 pg/μl for LM,which was consistent with conventional PCR.Conclusion The mLAMP described can potentially facilitate simultaneous detection of three kinds of bacteria in a large number of food samples, which could be used as a primary screening method and as a supplement to classical detection methods.
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The army research institutes and technical colleges bear the task of teaching,research and other functions,has a more comprehensive research support conditions and strong scientific and technological personnel,and undertake a large number of countries,the armed forces and local research projects,to obtain a large number of scientific and technological achievements,but how the equipment category results into productivity and combat effectiveness,technology management department,the problems to be solved.Scientific and technological achievements of military research institutes and technical colleges unconverted equipment class of 80,using analytic hierarchy process,a systematic analysis of the factors restricting equipment and technology achievements unconverted,mainly to solve the measures by questionnaire survey.
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Objective To study the serotype , biochemical characteristics , virulence gene and multilocus sequence typ-ing(MLST) of S.flexneri 4c in Beijing, Shanghai and Shenyang .Methods Seventy-six strains of S.flexneri 4c isolated from stool samples which had been collected from above-mentioned cities of China were identified with Denka Seiken serum and MASF monoclonal serum .Biochemical characteristics of each strain were identified by API 20E test strip and PCR technology was used for detecting 12 pair virulence genes of S.flexneri.MLST was used to analyze the characteristics . Results The serum agglutination antigen structure of S.flexneri 4c was(Ⅳ:7,8).MASF:B+,Ⅳ:Ⅰ+,7 (8) +.S.flexneri 4c developed different results in biochemical reactions and carried different rates of virulence genes , respectively .The IND test positive rate was 17.11%; MEL weakly positive rate was 3.9%, and ARA test weakly positive rate was 22.37%. Virulence genes were carried at a rate of 89.47% -100%, MLST typing was ST245.Conclusion S.flexneri 4c with serum agglutination antigen structure (Ⅳ:7,8) is a new serotype of S.flexneri.The main biochemical reactions are glucose fermentation and mannitol decomposition .A variety of Shigella related virulence genes are carried .MLST generation is consistent,suggesting that the bacteria might have evolved from ST 270 cloning.
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Objective To investigate the metabolism and function of the intestinal microbiota from liver cirrhosis patients.Methods Sixteen cases of liver cirrhosis and twenty normal individuals were selected , whose intestinal microbiota metagenomic DNA was extracted , followed by high-throughput Solexa sequencing and the bioinformatics analysis of metabo-lism and function annotation to compare the differences between the patients and normal subjects and find out about the cir -rhosis-related functions .Results The functional diversity was significantly reduced in the intestinal microbiota of cirrhotic patients.At the module or pathway level , the intestinal microbiota of patients showed an enrichment in metabolisms of drugs, essential amino acid , propanoate metabolism and inflammatory reaction , whereas an opposite tendency was observed in the metabolic ability of butyrate , bile acid and cell cycle .Conclusion Under the influence of liver cirrhosis , the growth environment in the intestine is destroyed , causing, the intestinal microbiota the exhibit some compensation to adapt to the changed intestinal micro-environment .
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Based on the work characteristics of disease and control of People's Liberation Army (PLA), the paper discusses the establishment of quality control system at Institute of Disease and Control of PLA and the subsequent effects. This could provide some reference for improvement disease control of PLA.
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DNA stable-isotope probing (DNA-SIP) is a recently developed method with which the incorporation of stable isotope from a labeled substrate is used to identify the function of microorganisms in the environment. The technique has now been used in conjunction with metagenomics to establish links between microbial identity and particular metabolic functions. The combination of DNA-SIP and metagenomics not only permits the detection of rare low-abundance species from metagenomic libraries but also facilitates the detection of novel enzymes and bioactive compounds. We summarize recent progress in SIP-metagenomic techniques and applications and discuss prospects for this combined approach in environmental microbiology and biotechnology.
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Animals , Humans , DNA , Genetics , DNA Probes , Chemistry , Genetics , Metabolism , DNA, Bacterial , Chemistry , Genetics , Metabolism , Isotope Labeling , Methods , Metagenomics , Methods , Molecular Probe Techniques , Sequence Analysis, DNA , MethodsABSTRACT
Objective To screen a human single-chain variable fragments(scFv)against antiGBM antibody.Methods Using phage display technique,the phage antibody library was panned by antiglomerular basement membrane(GBM)antibody which was coated in a micro-titer plate,one clone was found to have high affinity to anti-GBM antibody.The DNA sequence of the positive clone was determined.Results Along with the increase of rounds anti-GBM antibody specific phage antibody was highly enriched and screening efficiency was increased 137 folds than the firest round.ELISA and competition inhibition assay showed that the scFv had a specific combination character with anti-GBM antibody.DNA sequencing confirmed that the whole gene of scFv was 750 bp,and in accordance with humanized single-chain variable region antibody sequence structure.Conclusion The results suggested that the scFv fragment to anti-GBM antibody could be successfully selected by recombinant phage antibody technique,which will laid an experimental foundation for further research of the therapy of Goodpasture syndrome.
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Metaproteomics is an emerging proteomics technology to analyze large scale protein expression in environmental microbial ecosystem. It is termed as the large-scale characterization of the entire protein complement of environmental microbial community at a given point in time. This review focuses on the research strategies and the recent applications in this field based on the published reports and in combination with our own research experiences.
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Bacterial Physiological Phenomena , Ecology , Methods , Environmental Microbiology , Proteomics , MethodsABSTRACT
OBJECTIVE To detect ESBLs genes in 5 Shigella sonnei isolates.METHODS Susceptibility test to common antibiotics was performed through disk diffusion test,and ESBLs were confirmed according to CLSI.Conjugation experiment was performed to determine whether the resistance was transferable.The ESBLs gene was detected by PCR using universal primers for TEM,SHV,CTX-M,and the PCR products were also directly sequenced and analyzed.At the same time,the five isolates were analyzed by PFGE. RESULTS The five S.sonnei isolates were ESBLs producers and produced CTX-M-15 ESBLs,which were resistant to the most of the ?-lactams such as aztreonam,the first and second generation cephalosporins,cefotaxime,and non ?-lactam such as gentamicin,SF,but susceptible to ceftazidime,the 4th generation cephalosporins,Amikacin and meropenem.These strains were also intermediate to quinolones.CTX-M-15 gene could be transferred through conjugation.PFGE patterns of one isolate were different from the others.CONCLUSIONS Five S.sonnei isolates producing CTX-M-15 ESBLs are resistant to the most antibiotics.Clone spread is evident in these isolates.We should pay more attention to monitor these strains.
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Bacillus anthracis , the aetiological agent of anthrax, was the first discovered pathogenic bacterium in history Rapid progresses have been made on this field in recent years, especially; Bacillus anthracis has been sequenced successesfully early this year and published on the Internet The anthrax pathogenesis is always central to the study and there has been an enormous amount of work to elucidate it In this review, we will focus on the latest findings that concern three aspects of anthrax pathogenesis: Bacillus anthracis genome, pathogenic substances and pathogenesis mechanism
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In order to remove invasive plasmids from Shigella flexneri 2a 2457T and Shigella sonnei S7 based on the principle of plasmid incompatibility Ori and inc genes were amplified by PCR from S. flexneri 2a invasive plasmids, and then they were cloned into plasmid pMD18 T, resulting recombinant plasmids pMDori and pMDinc After S flexneri 2a 2457T and S sonnei S7 were transformed with pMDori or pMDinc respectively Invasive plasmids were removed from S flexneri 2a 2457T and S sonnei S7