Objective To investigate the alternations in gene/amino acid sequence of penicillin-binding protein (PBP)2x from clinical isolates of Streptococcus pneumoniae, and to find the reasons for the rapid surge of penicillin and cefotaxime nonsusceptibility among pneumococcal isolates from Shenyang. Methods Thirty-four strains of Streptococcus pneumoniae were collected from January 2006 to February 2007. The antibiotics susceptibility of these strains was detected. PCR amplification and direct sequencing of pbp2x genes were performed. The sequence variations of PBP genes of the penicillin nonsusceptible Streptococcus pneumoniae(PNSP) in this region were studied by BLAST analysis. Results Two prominent substitutions were common to 12 PNSP isolates for which the MIC of penicillin resistance and cefotaxime were at least 0.5 mg/L, which included the replacement of Thr338→Ala in the first conservative motif STMK and Leu546→Val adjacent to third conservative motif KSG. The importance of the exchange of His394→Leu was identified in one PNSP isolate 15. The remarkable finding in this study was Met342→Ile following the first conservative motif STMK. pbp2x sequences of eight PRSP isolates shared Lys501-Glu505-Thr507 substitutions which might be served as a unique marker for PRSP in this region. Novel gene and amino acid sequence variants in 17 isolate were identified in this study, and these sequences have been deposited in the GenBank database and assigned accession No. EU044831, EU089706-EU089709, EU106881-EU106884 and EU124672. Conclusion It is likely that the emergence of penicillin and cefotaxime nonsusceptible Streptococcus pneumoniae in Shenyang might be associated with novel gene sequence variants.

OBJECTIVE With surveillance of the distribution and antibiotic resistance of Escherichia coli during the last six years in our hospital,the basis for the reasonable clinical use of antibiotic is provided to doctor.METHODS A total of 1 907 strains of E.coli isolated during the last six years were analyzed by Kirby-Bauer disk or VITEK-2 system.RESULTS Among 1 907 strains of E.coli,1 114 strains were isolated from urine,accounted for 58.4%;215 from pus or secret,accounted for 11.3%;165 from sputum,accounted for 8.7%;and 159 from blood,accounted for 8.3%.ESBLs production rate of E.coli increased steadily from 5.11%,10.34%,14.56%,15.14%,33.79% to 29.96%,separately during the six years.The resistance of E.coli with ESBLs to most antibiotics was much higher than those without ESBLs.And E.coli demonstrated much higher resistant rate to ciprofloxacin,penicillins,and first or second generation cephalosporins,and much lower to amikacin and cefoperazone/sulbactam.No strains were found to be resistant to imipenem.CONCLUSIONS E.coli is the major pathogen,causing nosocomial infection with multi-resistant mechanism, since ESBLs-producing strain is increasing as years gone,reasonable choice of antibiotic should be in term of result of antibiotic resistant test and patient symptom to cure the E.coli infection induced.

OBJECTIVE To compare the changing trends of antibiograms of Salmonella typhi and S.paratyphi A isolates.METHODS A total of 42 isolates of Salmonella obtained from blood cultures patients in the First Affiliated Hospital of Chinese Medical University between 2001-2004 were included in the study.K-B test was used for the antimicrobial susceptivity test and the results were read based on National Committee for Clinical Laboratory Standards(NCCLS) of USA.RESULTS Twenty two isolates were identified as S.typhi and 20 were identified as S.paratyphi A.Sensitivity of S.typhi isolates to cephalosporins and chloramphenicol was found to have increased from 2001 to 2004 while that of S.paratyphi A showed a decline,meanwhile there was obviously increasing in resistance to ciprofloxacin.CONCLUSIONS Perhaps the schedule of empirical treatment of typhoid fever needs to be adjusted.

OBJECTIVE To study clinical distribution of Serratia and learn the antimicrobial susceptibility to S.marcescens in vitro in order to offer the reference to optimally selecting antibiotic.METHODS It was analyzed that the 222 Serratia strains were distributed in and the was deteted 164 S.marcescens strains were isolated from our hospital from 2001 to 2006.Their VITEK-2 of French Bio-M?rieux Company was adopted to proceed the identification of bacteria and antimicrobial susceptibility test.Data were analyzed by WHONET 5.4.RESULTS Serratia were mainly isolated from sputum,urine,blood,secretion,bile,cerebrospinal fluid,abdominal fluid,et al.Infection of both in-and out-patients could be caused by Serratia and most were in surgery ward.S.marcescens had higher drug resistance rates to piperacillin,cefazolin,cefuroxime,gentamicin and tobramycin which were all above 60%.They were all susceptible to imipenem(minimum inhibitory concentration only 1 ?g/ml) and their susceptible rates to piperacillin/tazobactam,ceftazidime,cefepime,and levofloxacin were all higher than 80%.CONCLUSIONS Serratia are less isolated from clinics,but have much higher antimicrobial resistance to the 1st and 2nd generation cephalosporin and show diversely drug resistance to 3rd cephalosporin,so physicians should pay attention to the infection caused by them.

Objective To investigate the alternations in gene/amino acid sequence of penicillin-binding protein (PBP) 2b from clinical isolates of penicillin-nonsusceptible Streptococcus pneumonia(PNSP) in this region.Methods 24 strains of Streptococcus pneumonia were collected from January to December 2006.The antibiotics susceptibility of these strains was detected.PCR amplification and direct sequencing of pbp2b genes were performed.The sequence variations of PBP genes of the PNSP in this region were studied with sequence BLAST analysis.Results Three prominent substitutions were common tO 13 PNSP isolates with minimal inhibitory concentration(MIC) at least 0.1 mg/L.These included the replacement of Thr445→Ala following the conservative motif SSN,Glu475→Gly and Thr488→Ala/Ser.The exchange of Glu332→Gly was identified in 12 PNSP isolates of which the MIC was at least 0.25 mg/L.Seven penicillin resistant Streptococcus pneumonia (PRSP) isolates (MIC≥3 mg/L)shared the amino acid substitution Ala618→Gly adiacent to third conserved (KTG) motif and the PBP2b sequences of seven PRSP isolates were classified within Back's group Ⅱ and were very similar to those of the Korean J77 isolate.Novel gene and amino acid sequence variants in isolate 14,15,8,11 and 24 was identified in this study and these gene sequences have been deposited in the GenBank database and assigned accession no.EU035970,EU056919,EU056920,EU056921 and EU106886.Conclusion Analysis of pbp2b genes revealed highly similar patterns of nucleotide and amino acid sequence variation among most resistant isolates.while penicillin intermediate Streptococcus pneumonia might be associated with novel gene sequence variants.

Objective To study the clinical significance of plasma(1→3)-β-D-Oaten measurement in invasive fungal infections.Methods The levels of plasma(1→3)-β-D-glucan were measured bymicrobiology kinetic rapid reader MB-80 and GKT-5M set dymmic fungus detecting kit in 14 patients proven to suffer from invasive fungal infection and 13 healthIy voluntary persons.And the difference between them was compared.Results In 14 patients with invasive fungal infection,8 patients had fungal infection of lower respiratory tract and lung,6 patients had fungemia.There were 11 patients infected by monilia(1 patient combined infection),2 patients infected by aspergillus,and 2 patients infected by pneumocystis(1 Datient clinical diagnosis without aetiology proof).The levels of plasma(1→3)-β-D-slucan in invasive fungal infections patients were(105.02±82.22)ng/L,which were higherthan thosein healthy persons[(6.65±1.01)ng/L)J,P＜0.01.Conclusion The levels of plasma(1→3)-β-D-glucanisan ia an important index in diagnosis of invasive fungal infections.

Objective To determine the possible genetic background and the source of our hospital's 43 clinical isolates of multidrug-resistant Acinetobacter baumannii, and the category of gene cassettes in type 1 integrons of all strains.Methods Restriction enzyme Apa I was chosed for all strains in pulsed-field gel electrophoresis (PFGE) methods.Multilocus sequence typing ( MLST) was used to compare the allelic profiles of all the strains. PCR method was used for amplify the integrons of all strains. Results PFGE results showed that 43 strains were divided into four types. A-type and B-type were divided into 4 and 2 subtypes, respectively. The MLST results showed the existing of three allelic profiles; 1-3-3-2-2-7-3, 1-3-3-2-2-11-3, and 1-3-3-2-2-14-3.B-type and D-type of PFGE have the same allelic profile(1-3-3-2-2-11-3).A-type strains were detected mainly in ICU, and in burn unit only found B- and D-type.The same integron was detected in 62.8% of the strains.The constituent ratio of A1,A2,A3,A4,B1,B2,C and D-type was 40.7% , 18.5% , 7.4% , 3.7% , 14.8% , 3.7% , 3.7% and 7.4% , respectively.Conclusions The coexistence of multiple cloning system in this region was proved by the PFGE and MLST, and the same clone can evolve to different subtypes when stimulated by different environmental conditions; and the different carrying-situationt of the same integron in strains prove the possibility of the change during the evolution of resistance mechanisms.

OBJECTIVE To offer the clinical physician the basis of optimal application of antibiotic,we have investigated the variation of antibiotic resistance and the bacterial spectra in the blood culture.METHODS Blood was cultured in BACTEC9120 of BD.The clinical isolates were identified by API and VITEK-2 of Bio-Merieux of France.Antibiotic susceptive test was done by Kirby-Bauer method and the result which was analyzed by WHONET5.3 and SPSS11.5 software was determined by the NCCLS standard of 2005′s edition.RESULTS Organisms were isolated from the blood specimen of 1468 patients,and there were 743 strains of Gram-positive cocci accounted for 50.7%,565 strains of Gram-negative bacilli accounted for 38.5%.Ninety three strains of fungi accounted for 6.3%.We analyzed the drug-susceptive result of Staphylococcus,Escherichia coli,and Klebsiella pneumoniae during five years,and found that all the antibacterial drug lost efficacy in some degree,except that the sensitivity of the staphylococci to vancomycin was 100%.CONCLUSIONS Gram-positive cocci are the main bacteria in blood culture,the species from which are diversified,and the rate of the drug resistance of some bacteria is high.It indicated that doctors should take more blood culture and monitor the bacteria drug resistant for the data of etiology,so that they can utilize antibiotic more reasonably.

OBJECTIVE To study the information of drug resistance of 1292 coagulase-negative staphylococci(CNS) isolates. METHODS There were 1292 isolates of CNS obtained from clinical patients in between 2001-2006.K-B test was used to determine the susceptibility and the results were read based on Clinical and Laboratory Standards Institute(CLSI) of the USA. RESULTS The isolating rate of CNS isolates was decreasing from 11.0% in the year of 2001 to 6.4% in the year of 2006.During the six years,the resistant rate to erythromycin,oxacillin and benzylpenicillin was high(90.0%,78.9% and 95.0%,respectively) and the resistant rate to cefepime,ciprofloxacin and levofloxacin increased from 17.3%,42.4%,and 28.7% in the year of 2001 to 39.1%,56.0% and 42.6% in the year of 2006.But the resistant rate to tetracycline,trimethoprim/sulfamethoxazole decreased from 71.2% and 72.0% in the year of 2001 to 52.7% and 33.3% in the year of 2006.The resistant rate to quinupristin /dalfopristin was 5.6%. CONCLUSIONS The steady resiatence rate of CNS over the past six years provides powerful evidence for the choice of clinical medication and practice of infection control.