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Objective:To analyze the clinical manifestations and test results of brucellosis cases, provide basis for clinical and laboratory differential diagnosis of brucellosis.Methods:Using retrospective analysis method, the cases of brucellosis confirmed by the Microbiology Laboratory of the Laboratory of Shenzhen People's Hospital from December 2015 to August 2019 were selected as the survey subjects, and the clinical data of the survey subjects were collected, including epidemiological characteristics, clinical manifestations, laboratory test results, treatment and prognosis.Results:Of the 14 patients, 9 were males and 5 were females. The median age of the patients was 42 years old (ranged 23 - 62 years old). Seventy-eight point six percent (11/14) of the patients were firstly diagnosed from April to September; 78.6% (11/14) of the patients had a history of contact with raw beef, mutton, pork and sheep placenta. The main clinical symptoms of brucellosis were fever (100.0%, 14/14) and arthralgia (42.9%, 6/14). The most common complication was arthritis (3/7). Laboratory results showed that the white blood cell count in all patients was not increased. However, many patients presented with high C-reactive protein level (9 cases), abnormal liver function (9 cases) and high erythrocyte sedimentation rate (8 cases). The median detection time of positive blood culture was 69 h (ranged 48 - 110 h). Seven patients had good prognosis, 4 patients continued to receive treatment due to complications, 2 patients failed to follow-up, and 1 patient died before the diagnosis was confirmed of the 14 patients.Conclusions:The clinical manifestations of brucellosis are complex, and detailed epidemiological history should be documented during clinical diagnosis to avoid misdiagnosis or missed diagnosis. Microbiologists should master the microbiological characteristics of Brucella, and improve differential diagnosis capabilities.
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Objective@#To investigate the prevalence of somatization symptoms and its correlation with social support in patients with depressive disorder.@*Methods@#Two hundred and fourteen hospitalized patients with depressive disorder were recruited. Patients were evaluated with Somatic Symptom Inventory (SSI), Perceived Social Support from Family Scale (PSS-Fa), Social Support Rating Scale (SSRS) and the general questionnaire.@*Results@#The average SSI scores of depressive patients was 49.63±15.53, with 26.2% (56/214) of the patients having moderate to severe level of somatic symptoms. The most common moderate to severe somatic symptoms in depressive patients were "feeling fatigued (61.3%, 131/214), weak (49.5%, 106/214), not feeling well (47.7%, 102/214), feeling faint or dizzy (48.6%, 104/214), or constipation (29.9%, 64/214)" . The average SSRS scores of patients was low (33.24±7.16). The SSI scores and its non-painful dimension (39.05±12.14) were significantly negatively correlated with family support (11.03±3.45) (r=-0.150, P=0.03; r=-0.153, P=0.02) .The SSI scores, and its somatization dimension scores (49.63±15.53) and painful dimension scores (10.58±4.18) were negatively correlated with the total scores of social support (33.24±7.16) (r=-0.164, P=0.02; r=-0.183, P=0.01; r=-0.136, P<0.05) and subjective support (19.14±4.81) (r=-0.167, P=0.02; r=-0.163, P=0.02; r=-0.177, P=0.01) . The SSI scores and its non-painful dimension of patients were negatively correlated with support utilization (r=-0.179, P=0.01; r=-0.194, P=0.01) .@*Conclusion@#Somatic symptoms are prevalent and severe in patients with depressive disorder. The somatic symptoms of depressive patients was closely correlated with decreased social support, especially low subjective support and support utilization.
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Objective To investigate the antimicrobial resistance profile of the clinical isolates collected from selected hospitals across China. Methods Twenty-nine general hospitals and five children's hospitals were involved in this program. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems. Results were interpreted according to CLSI 2017 breakpoints. Results A total of 190 610 clinical isolates were collected from January to December 2017, of which gram negative organisms accounted for 70.8％ (134 951/190 610) and gram positive cocci 29.2％ (55 649/190 610). The prevalence of methicillin-resistant strains was 35.3％ in S. aureus (MRSA) and 80.3％ in coagulase negative Staphylococcus (MRCNS) on average. MR strains showed much higher resistance rates to most of the other antimicrobial agents than MS strains. However, 91.6％ of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole, while 86.2％ of MRCNS strains were susceptible to rifampin. No staphylococcal strains were found resistant to vancomycin. E. faecalis strains showed much lower resistance rates to most of the drugs tested (except chloramphenicol) than E. faecium. Vancomycin-resistant Enterococcus (VRE) was identified in both E. faecalis and E. faecium. The identified VRE strains were mainly vanA, vanB or vanM type based on phenotype or genotype. The proportion of PSSP or PRSP strains in the non-meningitis S.pneumoniae strains isolated from children decreased but the proportion of PISP strains increased when compared to the data of 2016. Enterobacteriaceae strains were still highly susceptible to carbapenems. Overall, less than 10％ of these strains (excluding Klebsiella spp.) were resistant to carbapenems. The prevalence of imipenem-resistant K. pneumoniae increased from 3.0％ in 2005 to 20.9％ in 2017, and meropenem-resistant K. pneumoniae increased from 2.9％ in 2005 to 24.0％ in 2017, more than 8-fold increase. About 66.7％ and 69.3％ of Acinetobacter (A. baumannii accounts for 91.5％) strains were resistant to imipenem and meropenem, respectively. Compared with the data of year 2016, P. aeruginosa strains showed decreasing resistance rate to carbapenems. Conclusions Bacterial resistance is still on the rise. It is necessary to strengthen hospital infection control and stewardship of antimicrobial agents. The communication between laboratorians and clinicians should be further improved in addition to surveillance of bacterial resistance.
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Objective To analyze the pathogens distribution and drug resistance of nosocomial infection in patients with end-stage diabetic nephropathy .Methods 96 cases of end-stage diabetic nephropathy were randomly selected in our hospital .The speci-mens of urine ,blood and sputum were collected .The pathogens were identified by the drug susceptibility testing .Results The in-fection rate was 27 .08% .A total of 103 strains of pathogens were isolated ,including 15 strains of fungi ,42 strains of gram-negative bacteria and 46 strains of gram-positive bacteria .The drugs susceptibility rates of the fungi to flucytosine ,amphotericin B and flu-conazole were 100% ,and it also showed that the fungi had higher sensitivity to other common antibiotics ;the drug susceptibility rates of gram-positive bacteria to vancomycin and teicoplanin were 100% ,while its drug susceptibility ability to penicillin ,strepto-mycin and others were weak ;the drug susceptibility rates of gram-negative bacteria to piperacillin and imipenem were high ,while its drug susceptibility ability to aztreonam and cephalosporin were weak .Conclusion The low resistance in the patients with end-stage diabetic nephropathy the abuse of clinical antiseptic drugs lead to the dysbacteriosis ,resulting in a significant increase of the inci-dence of nosocomial infection ,so the analysis of pathogens distribution and drug resistance of nosocomial infection has clinical sig-nificance .
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Objective To investigate the prevalence and molecular characteristics of the extended -spectrum β-lactamase ( ESBL) and AmpC enzyme-producing Proteus mirabilis ( P.mirabilis) strains isola-ted in Shenzhen People′s Hospital.Methods The production of ESBLs and AmpC enzymes by P.mirabilis isolates were detected by a screening and confirmatory test for ESBLs and AmpC disk test , respectively .The PCR assays followed by DNA sequencing of the products were employed to analyze the multiple genes inclu -ding the ESBLs genes, AmpC genes, insertion sequences (ISs) upstream of the ESBLs or AmpC genes, plasmid -mediated quinolone resistance ( PMQR ) determinants , quinolone resistance-determining region (QRDR) genes , the integrase genes, and class1 integron cassette.The epidemiological analysis of the iso-lates was performed by pulsed field gel electrophoresis .Results There were 130 P.mirabilis clinical iso-lates collected from Shenzhen People′s Hospital in China during the year 2004 to 2010.Among them, 13 isolates (10%) produced ESBLs, that accounted for 0%-9.1%in the year 2004-2009 and up to 29.4%in 2010, and 3 isolates (2.3%) produced AmpC enzymes.The predominant genotype of ESBLs -producing isolateswas b al CTX-M-14(n=7), followed by blaCTX-M-65(n=3), blaCTX-M-55(n=1), blaCTX-M-24(n=1) and blaPER-1 (n =1).The clinical isolate of PER-1-producing P.mirabilis was reported for the first time in China.Twoisolates carried an AmpC β-lactamase gene of blaCMY-2 and one isolate carried an unidentified AmpC gene .ISEcp1 located upstream of blaCTX-M and blaCMY-2 were detected in 91.7% (11/12) of CTX-M-producing isolatesand one CMY-2-producing isolate, respectively.ISPa12 was present upstream of blaPER-1 in one studiedisolate.Approximately 66.7% (10/15) of ESBL and /or AmpC-producing isolates harbored PMQR genes including2 carrying qnrD, 5 carrying aac-Ib-cr and 3 carrying both qnrD and aac-Ib-cr.Twelve ESBL and /orAmpC-producers with high level of resistance to ciprofloxacin carried the similar mutation profiles of S 83I inGyrA, S80I or S80R in ParC and among them, six strains showed E466D mutation in GyrB.Approximately86.7% (13/15) of ESBL and/or AmpC-producing isolates carried class 1 integron.Fourteen PFGE typeswere observed among 15 ESBL and/or AmpC-producers.Conclusion The prevalence of CTX-M β-lactamasesin P.mirabilis isolates contributed to the increased resistance to extended -spectrum cephalosporins.The qnrD and/or aac-Ib-cr genes were detected among the most of ESBL and /or AmpC-producing P.mirabilis clinical isolates.
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Objective To evaluate the capability of multilocus variable-number tandem-repeat ( VNTR) analysis ( MLVA) and pulsed-field gel electrophoresis ( PFGE) for genotyping Salmonella enterica serovar Typhi (S.Typhi) isolates.Methods Five polymorphic VNTRs (SAL02,SAL11,SAL16,SAL20, and TR4699 ) that were observed in S.Typhi strains from previous studies were selected to establish MLVA . Twenty-one epidemiologically unrelated S.Typhi strains that were isolated from Shenzhen ,China from 2002 to 2007 were genotyped by the established MLVA , and the results were compared with those by PFGE . Results The Simpson′s index of diversity ( D value ) for all five different VNTRs ranged from 0.838 to 0.943 .A total of 19 MLVA profiles and 19 PFGE profiles were found , respectively .The D value for both MLVA and PFGE were 0.99 and the test results from two analyses were identical .Conclusion The five polymorphic VNTRs analysis could be used as an alternative typing scheme for epidemiologic investigation of S.Typhi infection .
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Objective To study the epidemiological features of Cryptococcus neoformans and Cryptococcus gattii isolated from clinical samples in Shenzhen and to elucidate the distribution of species,varieties,genotypes and mating types within the strains tested.Methods The strains involved in this study were 55 cryptococcal strains isolated from our clinical samples.The canavanine-glycine bromthymolblue (CGB) culture was performed to distinguish Cryptococcus neoformans from Cryptococcus gattii.The genotype was characterized by polymerase chain reaction (PCR) fingerprinting with primer M13.The Cryptococcus gattii species and varieties of grubii and neoformans together with two opposite mating type α and a were identified by PCR with variety-specific and mating type-specific primers.The GEF1-restriction fragment length polymorphism analysis was conducted to simultaneously determine the genotype and mating types of strains tested.The sequence type of IGS1 region was analyzed for the VG Ⅱ genotype.Results Of the 55 tested cryptococcal strains,52 were Cryptococcus neoformans,all of which were var.grubii,genotype VN Ⅰ and mating type α.The remaining 3 strains were Cryptococcus gattii,among which,one was genotype VG Ⅰ and mating type α,and two were genotype VG Ⅱ and mating type α.The two VGⅡ genotype strains belonged to the sequence type Ⅱ.Conclusions The strains belonging to the Cryptococcus neoformans var.grubii,genotype VN Ⅰ and mating type α predominate in causative pathogens of cryptococcosis in Shenzhen.Cryptococcus gattii accounts for minority of the cryptococcal isolates,and the highly pathogenic VG Ⅱ genotypes in foreign countries are also characterized.The sequence types of IGS1 region of the two VG Ⅱ strains are in accord with VG Ⅱb sub-genotype.
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Objective To investigate the antimicrobial resistance mechanisms and genetic homogeny of Salmonella from community acquired infections in Shenzhen,China.Methods Ninety-three of Salmonella were isolated from 2002 to 2007 at Shenzhen People's Hospital,China.PCR and DNA sequencing were used to investigate the mutation in QRDR of the gyrA,gyrB,parC and parE.Plasmid mediated quinolone resistance genes including qnr and aac(6')-Ib-cr,β-lactamase genes including blaTEM,blaSHV,blaOXA, blaCTX-M, and class 1 integron were detected. All isolates were typed by PFGE. Results S. enterica typhi and S. enterica paratyphi A were susceptible to ampicillin, chloramphenicol, trimethoprim/sulfamethoxazole, ceftriaxone and ciprofloxacin, with the susceptible rate of 96%-100%. Fifty-two percent (13/25) of S. enterica typhi and 95% (61/64) of S. enterica paratyphi A were resistant to nalidixic acid. Twenty-four percent (6/25) of nalidixic acid-resistant S. enterica typhi and 94% (60/64) of nalidixic acid-resistant S. enterica paratyphi A showed decreased susceptibility to ciprofloxacin (MIC of 0. 125-1 μg/ml).All nalidixic acid-resistant (susceptible to ciprofloxacin ) Salmonella (NARS) isolates had a single substitution in the QRDR of GyrA, and 91% (68/75) of these isolates carried the substitution Ser83Phe in GyrA. Two mutations in the QRDR of GyrA were detected in both of two ciprnfloxacin-resistant Salmonella,with the additional one mutation in the QRDR of parC. Plasmid mediated quinolone resistance genes including qnr and aac(6')-lb-cr were not detected in any isolate. The blaCTX-M-14 gene was detected in a ceftriaxoneresistant isolate of S. enterica paratyphi A, with ISEcpl located on the upstream of it. Three muhidrugresistant strains of Salmonella all carried one 1 900 bp classⅠ integron gene cassette dhfrⅫ-orfF-aadA2,with the additional one β-lactamase gene of blaTEM-1, or blaOXA-30. Twenty-two distinct PFGE patterns were observed among twenty-five S. enterica typhi. The PFGE patterns of sixty-four S. enterica paratyphi A showed limited genetic diversity (average similarity of 91% ). Ninety investigated inpatients were infected in the community. Six patients infected by S. enterica paratyphi A had a travel history before infection. Conclusions Nalidixic acid-resistant S. enterica typhi and S. enterica paratyphi A are highly prevalent in Shenzhen,China. The mutation in the QRDR of GyrA is the prevalent mechanism responsible for the resistance to nalidixic acid in Slmonella. The great genetic similarity among S. enterica paratyphi A isolates indicates endemic disease from the presence of a single clone over 6-year period.
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In order to express the gene of LEN-5 β-lactamase from a Klebsiella pneumoniae strain,plasmids in the strain were extracted and an 879bp product of LEN-5 gene was obtained with PCR.After being digested with Nde I and Xho I,LEN-5 gene was cloned into pET-26b (+) vector.Then it was confirmed by digestion and DNA sequencing in recombinant plasmid before transformed into E.coli BL21 (DE3).After inducing by IPTG,LEN-5 β-lactamase was expressed.Protein extraction was processed by ultrasonic and protein activity was detected by nitrocefin.The isoelectric focusing electrophoresis showed a pI of 7.6.These results indicated that the LEN-5 gene has been cloned and expressed in prokaryote cell successfully.
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OBJECTIVE To investigate the prevalence of multidrug resistance(MDR) mechanisms of Staphylococcus haemolyticus against oxacillin,gentamycin and erythromycin.METHODS Agar dilution method was performed to detect the minimal inhibition concentration(MIC) of 3 antimicrobial agents against 63 strains of S.haemolyticus,and the resistance genes of mecA,aac(6′)+aph(2″),ermA,ermB,ermC and msrA/msrB were investigated by PCR in all clinical isolates.RESULTS mecA Gene was detected in 62 isolates of meticillin-resistant S.haemolyticus(MRSH),and aac(6′)+aph(2″) gene was found in 50 isolates resistant to gentamicin,and the most prevalence erythromycin resistance gene in S.haemolyticus was msrA/msrB(58.7%),followed by ermC(31.7%).Among the 43 MDR strains,the more commonly encountered three genes were mecA,aac(6′)+aph(2″) and msrA/msrB(58.1%)or ermC(20.9%),and 8 isolates(18.6%) were found harboring four genes of mecA,aac(6′)+aph(2″),ermC and msrA/msrB.CONCLUSIONS The mecA,aac(6′)+aph(2″),msrA/msrB and ermC genes are main resistance mechanisms against oxacillin,gentamicin and erythromycin in mutidrug resistant S.haemolyticus.
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OBJECTIVE To investigate the antimicrobial susceptibility of 9 antimicrobial agents and multidrug(resistance)(MDR) phenotype among Acinetobacter baumannii isolated from hospitalized patients.METHODS Disk diffusion method was used to detect the inhibitory zone diameter of 9(antimicrobial) agents to 221 strains of A.baumannii.according to standard from NCCLS in South China during 2002 to 2004.RESULTS 25.3% and 26.4% of isolates from ICU patients showed resistance to(imipemem) and meropemem,respectively,and the resistance rates were greater among the(isolates) from ICU(patients) than those from non-ICU inpatients by 13.4% and 12.2 %(P
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OBJECTIVE To investigate the susceptibilities of Staphylococcus aureus and Enterococcus isolated from Shenzhen Hospital during 2005 to 2006 to 17 antimicrobial agents.METHODS The minimal inhibitory concentrations(MICs) of 17 antibacterial agents were determined by agar dilution method,WHONET5.3 software was used to analyze the data.RESULTS The prevalence of meticillin-resistant S.aureus(MRSA) was 20.7%.Vancomycin and teicoplanin remained very high activity against MRSA(100%),with the MIC50 and MIC90 of 0.5 and 1 ?g/ml,respectively.Fluoroquinolones(ciprofloxacin,levofloxacin and gatifloxacin),trimethoprim/sulfamethoxazole,erythromycin,and clindamycin showed very low activity against MRSA(0-33.3%).The most active agent against Enterococcus faecalis was vancomycin and teicoplanin(100.0%),followed by piperacillin/tazobactam,imipenem and ampicillin(97.2-100.0%).E.faecium showed high resistance to various agents,except to vancomycin and teicoplanin(susceptible rate 100.0%).Cross-resistance to fluoroquinolones was found among MRSA,MSSA,E.faecalis,and E.faecium.CONCLUSIONS MRSA and E.faecium isolated from our hospital showed high resistance to multiple antimicrobial agents except vancomycin and teicoplanin.