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Objective:To investigate the serotype distribution, drug resistance and the virulence gene profiles of clinical isolates of Salmonella in Zhejiang Province. Methods:A retrospective study of 463 clinical isolates of Salmonella in nine regions of Zhejiang Province from 2010 to 2017 was conducted. Their serotypes were detected using the Kauffmann-White scheme based on serological detection. Antibiotics susceptibility tests were carried out using K-B disk diffusion method. Eleven virulence genes were detected by polymerase chain reaction. Results:A total of 35 serotypes were identified among the 463 strains of Salmonella. The dominant serotypes were Salmonella typhimurium (41.90%(194/463)) and Salmonella enteritis (22.25%(103/463)). The resistance rate of Salmonella to ampicillin was the highest (66.52%(308/463)), followed by ampicillin/sulbactam (46.87%(217/463)), while low resistance rate to piperacillin/tazobactam was observed (3.24%(15/463)). All strains were sensitive to imipenem and meropenem. Finally, 188 strains (40.60%) of multi-drug resistance were found. The virulence genes hilA, ssrB, marT, siiD, sopB and pagN were found in all Salmonella strains. The virulence gene vexA was found only in Salmonella typhi and Salmonella Dublin; virulence gene icmF was mainly found in Salmonella enteritis. The virulence plasmid gene spvB and pefA were mainly found in Salmonella enteritidis and Salmonella typhimurium and invariably appeared in pairs. The virulence gene cdtB was mainly found in Salmonella typhi and Salmonella paratyphi A. Conclusions:Salmonella typhimurium and Salmonella enteritis are the main clinically isolated Salmonella strains in Zhejiang Province. The situation of multi-drug resistance is severe and a variety of virulence genes are highly detected.
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Objective To study the clinical effect of Yaotongning capsule combined with etoricoxib for the pain and inflammation of lumbar vertebrae in elderly patients with lumbar osteoarthritis. Methods 120 elderly patients with lumbar osteoarthritis admitted to our hospital from January 2016 to June 2018 were randomly divided into the control group and the observation group, with 60 patients in each group. Patients in the control group were treated with etoricoxib, while patients in the observation group were treated with etoricoxib plus Yaotongning capsule orally. Both groups received medications for 2 weeks. Spinal pain and quality of life score changes were recorded. The inflammatory cytokines in serum TNF-α, GM-CSF, COX-2 and BMP-2 levels were monitored. The clinical efficacy was compared and drug safety profile was evaluated for two groups. Results The effective rates of the control group and the observation group were 78.33% and 91.67% respectively. The effective rate in the observation group weas significantly higher (P<0.05). After treatment, the VAS score for the patients in the observation group was significantly lower than that in the control group (P<0.05). The SF-36 score in the observation group was significantly increased (P<0.05), and the levels of TNF-α,GM-CSF and COX-2 in the serum were significantly lower than those in the control group (P<0.05), and the levels of BMP-2 were significantly increased (P<0.05). Conclusion Yaotongning capsule combined with etoricoxib in the treatment of senile lumbar osteoarthritis has definite curative effect. It significantly reduced lumbar pain, improved quality of life, inhibited inflammatory reaction, and had a better drug safety profile. The further clinical investigation for the combination therapy is warranted.
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Objective To analyze the sequence of Pre-S gene in asymptomatic chronic HBV carriers (ASCs) with low-level HBsAg.Methods The serum samples were collected from 654 ASCs in the First Affiliated Hospital of Zhejiang University School of Medicine , Hangzhou Sixth People’s Hospital and the 903th Hospital of PLA.According to the level of HBsAg , ASCs were divided into low-level HBsAg group (≤10 IU／mL ) and high-level HBsAg group (＞10 IU／mL ).The pre-S／S gene amplification and sequencing were performed in 138 ASCs with low-level HBsAg and 100 age-matched ASCs with high-level HBsAg.A phylogenetic tree was constructed to determine the genotype , based on the successful sequencing results of Pre-S gene in the high level HBsAg group , the Pre-S gene reference sequences of the main ASCs genotypes in Eastern China were established.The sequence of Pre-S gene in low-level HBsAg group was analyzed and compared with the reference sequences.SPSS 12.01 statistical software was used to analyze the data.Results Sixty-three cases of Pre-S／S were successfully sequenced in 138 ASCs of low-level HBsAg group, including 52 cases of B genotype and 11 cases of C genotype.Among the 100 cases of high-level HBsAg group, 94 cases of Pre-S／S were successfully sequenced , including 48 cases of B genotype and 46 cases of C genotype.The sequence analysis indicated that in the B genotype , 81 amino acid mutation sites were found in the Pre-S protein of the low-level HBsAg group, including 4 significant mutations: F56I／V, T76A／N／P in the Pre-S1 region, P15L／S／T and Y21T／F／H／N in the Pre-S2 region; while 47 amino acid mutation sites were found in Pre-S protein of high-level HBsAg group, including 3 significant mutations :L34F, V49A and P59S／L in Pre-S1 region.The total number of amino acid mutation sites in the low-level HBsAg group of B genotype was higher than that of the high-level HBsAg group (χ2 ＝14.008, P＜0.05). In the C genotype, 19 amino acid mutation sites were found in the Pre-S protein of the low-level HBsAg group, including 3 significant mutations : W66V／G and A79V in the Pre-S1 region,V32A in the Pre-S2 region; while 39 amino acid mutation sites were found in Pre-S protein of the high-level HBsAg group, including 2 significant mutations: A79V in Pre-S1 region and T49I in Pre-S2 region.The total number of amino acid mutation sites of Pre-S protein in the C genotype was significantly different between the two groups (χ2 ＝7.571, P＜0.05).Conclusion Significant mutations in Pre-S gene may be associated with the persistent expression of low-level HBsAg in ASCs.
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With the abuse of antimicrobial agents in developing countries, increasing number of carbapenem-resistant Enterobacteriaceae (CRE) attracted considerable public concern. A retrospective study was conducted based on 242 CRE strains from a tertiary hospital in Hangzhou, China to investigate prevalence and drug resistance characteristics of CRE in southeast China. Bacterial species were identified. Antimicrobial susceptibility was examined by broth microdilution method or epsilometer test. Resistant β-lactamase genes were identified by polymerase chain reaction and sequencing. Genotypes were investigated by phylogenetic analysis. Klebsiella pneumoniae and Escherichia coli were the most prevalent types of species, with occurrence in 71.9% and 21.9% of the strains, respectively. All strains exhibited high resistance (> 70%) against β-lactam antibiotics, ciprofloxacin, trimethoprim-sulfamethoxazole, and nitrofurantoin but exhibited low resistance against tigecycline (0.8%) and minocycline (8.3%). A total of 123 strains harbored more than two kinds of β-lactamase genes. bla, bla, bla, and bla were the predominant genotypes, with detection rates of 60.3%, 61.6%, 43.4%, and 16.5%, respectively, and were highly identical with reference sequences in different countries, indicating potential horizontal dissemination. IMP-4 was the most frequent class B metallo-lactamases in this study. In conclusion, continuous surveillance and effective prevention should be emphasized to reduce spread of CRE.
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Humans , Anti-Bacterial Agents , Therapeutic Uses , Carbapenem-Resistant Enterobacteriaceae , Genetics , China , Epidemiology , Enterobacteriaceae Infections , Epidemiology , Microbiology , Genotype , Microbial Sensitivity Tests , Phylogeny , Prevalence , Retrospective Studies , beta-Lactam Resistance , beta-Lactamases , GeneticsABSTRACT
Objective To evaluate the reliability of the inhibitor enhanced carbapenem inactivation method (ieCIM) in the detection and preliminary classification of carbapenemase in gram-negative rods.Methods The carbapenem inactivation method (CIM) was modified by adding tazobactam or ethylene diamine tetraacetic acid disodium salt as carbapenemase inhibitors into the reaction system.A total of 198 isolates of Enterobacteriaceae and 35 strains of nonfermenters were collected,and their preliminary classification of carbapenemase was performed by the ieCIM.Meanwhile,their carbapenemase genes were detected by polymerase chain reaction (PCR),and the results were compared with that of the ieCIM.Results Among 198 strains of Enterobacteriaceae,101 were positive for carbapenemase genes,while 99 were detected by the CIM.Among the other 97 strains with negative carbapenemase gene,the results of the ieCIM were also negative.Among 35 strains of nonfermenters,25 were positive for carbapenemase genes,while 24 were detected by the CIM.Among the other 10 strains with negative carbapenemase gene,the results of the CIM were also negative.Using the ieCIM,97.7％ (85/87) of strains producing class A carbapenemase and 88.0％ (22/25) of strains producing class B carbapenemase were detected.Twelve strains producing class D carbapenemase and 2 strains producing both class A and class B carbapenemase were detected by the ieCIM.The total detection sensitivity and specificity of the ieCIM were 96％ and 100％,respectively.Conclusion The ieCIM has the consistent results with the detection method of carbapenemase genes,which may be used to detect and classify carbapenemase in clinical microbiology laboratories.
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In recent years , respiratory tract infectious diseases and public health events caused by human adenovirus type 7(HADV-7) had been rising.Especially in Asia, the outbreaks of acute respiratory illness caused by HADV-7, accounting for more than 60%in the world, had brought great influence to the public health.In order to prevent the occurrence of HADV-7 epidemic and improve the diagnostic performance of laboratory , the research progress of HADV biological features and typing , HADV-7 epidemiological characteristics , clinical and differential diagnosis and laboratory examination methodologies were reviewed in this paper.
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Objective@#To investigate the association between hepatic controlled attenuation parameter (CAP) and metabolic syndrome (MetS) and the correlation of CAP and its changes with the incidence of MetS.@*Methods@#A total of 2461 subjects who underwent physical examination from July 2013 to September 2015 were enrolled. Spearman correlation analysis was used to investigate the correlation of CAP with the number of MetS components and each MetS component, and the chi-square test was used to investigate the prevalence rates of MetS and each component under different CAP levels. Logistic regression analysis was used to analyze the odds ratio (95% confidence interval (CI)) of MetS under different CAP levels. A total of 230 subjects without baseline MetS were selected; in a prospective cohort study, these subjects were divided into groups according to the baseline CAP, change in CAP, and percent change in CAP, and the chi-square test was performed to compare the incidence of MetS. The Cox regression analysis was used to analyze the values of baseline CAP, change in CAP, and percent change in CAP in predicting MetS.@*Results@#CAP was positively correlated with the number of MetS components (r = 0.309, P < 0.01) and significantly correlated with all components. There were significant differences in the prevalence rates of MetS and its components under different CAP levels (< 238 dB/m, 238-258 dB/m, 259-291 dB/m, and ≥292 dB/m) (P < 0.05). After the adjustment for sex and age, with < 238 dB/m as a reference, the odds ratios (95% CI) of MetS in patients with CAP levels of 238-258 dB/m, 259-291 dB/m, and ≥292 dB/m were 1.784 (1.369-2.325), 2.936 (2.292-3.760), and 4.363 (3.435-5.543), respectively (all P < 0.05). Follow-up data showed that 28 patients (12.2%) developed MetS. After the adjustment for related factors, the hazard ratios (95% CI) of MetS in patients with baseline CAP > 238 dB/m, change in CAP > 30 dB/m, and percent change in CAP > 25.0% were 3.337 (1.163-9.569), 7.732 (2.453-24.366), and 11.656 (3.329-40.813), respectively (all P < 0.05).@*Conclusion@#CAP is closely associated with MetS and its components. CAP and its change can be used to predict the risk of MetS.
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Objective To evaluate the performance of Carba NP test (CNP) for detection of carbapenemases in carbapenem-non-susceptive Enterobacteriaceae.Methods Carbapenemases in Enterobacteriaceae strains isolated from clinical specimens in the 117th hospital of PLA from January 2010 to December 2014 were detected by both CNP and modified Hodge test (MHT),and corresponding results were analyzed and compared by Chi-square using SPSS version 19.0.Data acquired from PCR were used as the standard.Results Of the 253 carbapenem-non-susceptive Enterobacteriaceae strains,188 positive strains and 65 negative strains were detected by MHT,and 175 positive strains,77 negative strains and 1 undetectable strain were detected by CNP,while 177 positive strains and 76 negative strains were detected by PCR.Sensitivities of two tests were comparable [CNP,98.9％ (175/177),versus MHT,98.3％ (174/177);x2 =0.5,P =0.48],but CNP was more specific [100.0％ (76/76) versus 81.6％ (62/76);x2 =12.1,P ＜ 0.05].Positive result in the CNP was emerged earliest in a KPC-producing strain.Conclusion The CNP is rapid and simple,which have good sensitivity and specificity.Yet further perfection is still needed in the future.
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Objective To evaluate inhibitory effects of the Chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia psittaci strain guinea pig inclusion conjunctivitis (GPIC) and Chlamydia trachomatis (Ct) serovar E,and to provide new ideas for the treatment of Ct infection.Methods The Chlamydiaphage phiCPG1 capsid protein Vp1 was expressed in Escherichia coli BL21 transfected with the recombinant plasmid Vp1-pET30a (+),identified by Western blot analysis and purified by using dialysis bags.Bicinchonininc acid (BCA) assay was performed to determine the concentration of Vp1 protein.GPIC and Ct serovar E strains were both classified into 4 groups to be firstly incubated with Vp1 protein (Vp1 group),Tris-glycine solution (Tris group),S protein (S group) or Dulbecco's Modified Eagle Medium (DMEM,DMEM group) at room temperature for 3 hours,then were used to infect Hela cells followed by 72-hour (GPIC) or 48-hour (Ct serovar E) culture with the presence of Vp 1 protein (Vp 1 group),Tris-glycine solution (Tris group),S protein (S group) or DMEM (DMEM group).Subsequently,immunofluorescence staining was conducted to observe and count chlamydial inclusions.Results The number of GPIC inclusions was significantly different between the 4 groups after 72-hour culture (F=476.632,P＜ 0.05),and lower in the Vp1 group (5.0 ± 1.5) than in the Tris group (24 ± 1.2,P＜ 0.05),S group (25 ± 1.7,P＜ 0.05) and DMEM group (25 ± 1.5,P＜ 0.05),but insignificantly different between the latter 3 groups (P ＞ 0.05).Compared with the DMEM group,the Vp1 group showed a significant decrease of 80.2％ ± 3.99％ and 77.2％ ± 1.79％ in the number of GPIC and Ct serovar E inclusions respectively,with no significant difference in the inhibitory effect of Vp1 on GPIC versus Ct serovar E (t =2.057,P ＞ 0.05).Conclusion The phiCPG1 capsid protein Vp1 can obviously inhibit GPIC and Ct serovar E infections to a similar degree.
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Applications of molecular diagnostics in clinical microbiology laboratory were introduced in the article,including nucleic acid hybridization,nucleic acid amplification,DNA sequencing,gene chips and mass spectrometry.Molecular diagnostic techniques provide major tools for rapid diagnosis of infectious diseases,molecular epidemiology investigation,rapid identification of microbial pathogens and the study of the pathogenicity and antibiotics resistance.The application of these new technologies,as the complement of traditional culture methods,increases sensitivity,accuracy and diagnostic efficiency of assays.
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Major changes and updates of Clinical and Laboratory Standards Institute (CLSI) document M100-S22 for performance standards for antimicrobial susceptibility testing were introduced in the article,which includes ( 1 ) general changes; (2) changes of drugs recommended for testing and reporting;( 3 ) changes of interpretive criteria ( breakpoints ) and comments ; ( 4 ) changes of quality control and others; and (5) changes of appendixes and glossaries.
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Recent major changes and updates of Clinical and Laboratory Standards Institute(CLSI) document M100 for performance standards for antimicrobial susceptibility testing were introduced in the article,which include changes of interpretive criteria and comments,antimicrobial susceptibility testing of infrequently isolated or fastidious bacteria,changes of appendixes and others,Brief comments were made for these changes.
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ObjectiveTo investigate correlation between drug-resistance related genes and mobile genetic elements of Klebsiella pneumoniae resistant to β-lactams. Methods Forty-seven strains of multidrug-resistant Klebsiella pneumoniae were collected from 6 hospitals in Hangzhou and Huzhou of Zhejiang province from August 2008 to May 2010.Modified Hodge test was performed to detect phenotypes of carbapenemases.Forty kinds of β-lactamases (class A-D),ompK35,ompK36,and 12 kinds of mobile genetic elements were detected by PCR,and the results were analyzed by index cluster.ResultsThirty-five strains were positive in modified Hodge test,and 5 kinds of β-lactamases gene ( including KPC-2-like,GenBank:HQ258934) and 9 kinds of mobile genetic elements were detected.Mutations were observed in ompK35 and ompK36 when compared with sensitive strains.Index cluster analysis showed that correlation existed between KPC-2,KPC-2-like and ISKpn6,between TEM-1 and ISEcpl,IS26,int Ⅰ 1,trbC,IS903,and between CMY-2,OXA-30,DHA-1 and tnpU,tnp513,trbC.ConclusionsFive kinds of β-1actamases genes,and mutations in ompK35 and ompK36 may be associated with the resistance to β-1actams in multidrug-resistant Klebsiella pneumoniae.
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Objective To investigate the molecular characteristics and epidemiological signification of patients with low-level HBsAg. Methods PCR and gene sequencing were used to detect HBV DNA and Tyr-Met-Asp-Asp(YMDD) mutant in 136 serum samples with low-level HBsAg and 44 sernm samples with high-level HBsAg. Genotyping was performed in 47 cases with HBV DNA 10~5 copies/L by concentration method and 37 cases with high-level HBsAg. S gene sequences and serotypes were analyzed in 14 cases with HBV DNA 105 copies/L and 29 cases with high-level HBsAg. S gene sequences were compared with the consensus sequence of Chinese strain by BioEdit software. Results The HBV DNA-positive rate, YMDD mutation rate and HBV DNA load (logarithm) in low-level and high-level HBsAg group were 34.6% (47/136), 0% (0/136), 6.5±1.4 and 84.1% (37/44), 9.1% (4/44), 8.9±1.8, respectively. There was statistically significant differences between two groups (for concentration method,χ~2 = 30.8, P < 0.05; for direct method, χ~2 = 53.5, P < 0.05; for YMDD mutation ratio, P = 0.003, For HBV DNA (log), t = 6.5, P < 0.05). The genotypes in low-level HBsAg group included type B (16/47), type C (5/47) and non-classified ones(26/47). There were significant differences between two groups (χ~2=21.8, P <0.01). The serotypss included adw (7/14), ayw (4/14), adr (2/14) and ayr (1/14). There were significant differences in genotypes (χ~2 = 13.5, P < 0.05) but not in serotypes between two groups (χ~2 = 4.7, P >0.05). S gene sequencing results showed no S gnne variation was detected, but there were 6 single nucleotide polymorphisms in 16 cases, which would not result in the alternation of amino acid. Conclusions Low-replication phenomenon of HBV DNA was present in patients with low-level HBsAg. The major genotyps and serotype was type B and adw/ayw, respectively. Polymorphic variants have been found in the S gene. The existence of low-level HBsAg might be related with its own molecular characteristics resulting in low expression of HBsAg or immune tolerance induced by low-level HBsAg after HBV infection.
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OBJECTIVE To investigate the distribution and the drug resistance of Klebsiella pneumoniae in our hospital,and provide reference for the rational use of antimicrobial agents in clinic.METHODS The strains of K.pneumoniae isolated from clinical samples were identified by the VITEK-32 system.Extended spectrum ?-lactamases(ESBLs) were determined by phenotypic confirmatory test.Susceptibility test to antimicrobial agents was performed by disk diffusion methods and analyzed by WHONET 5.3.RESULTS 93.6% Of total 256 strains of K.pneumoniae were isolated from urine,sputum,blood and secretion samples,and 54.3% of them were isolated from sputum.81.6% Of infections caused by K.pneumoniae were frequently occurred in patients in intensive care units(ICU),departments of respiratory medicine and gastroenterology,departments of hematology and endocrinology,cadred wards and department of nephrology.The isolated ratio of ESBLs producing K.pneumoniae were 39.8%.The rates of resistance to piperacillin,cefuroxime,ceftazidime, cefotaxime,ceftriaxone,aztreonam,gentamicin,ciprofloxacin,and sulfamethoxazole/trimethoprim were between 32.4% and 50% in all strains;and the resistance rate to cefoperazone/sulbactam,piperacillin/tazobactam,cefepime,cefoxitin,and amikacin was less than 22.7%.There were isolates producing ESBLs,which showed much higher resistance to antimicrobial agents tested(but susceptible to imipenem and meropenem) than that of non-ESBLs-producing K. pneumoniae(P