Comparison of the results of disc diffusion methods applied for detection of ESBL-positive strains of gram-negative rods

Examinations were undertaken to compare the results of disc diffusion tests applied for detection of strains producing extended-spectrum beta-lactamases (ESBLs). A total of 120 clinical strains were used in experiments. These strains were determined as ESBL-positive on the basis of consistent results of two methods: the double disc synergy test (DDST) according to Jarlier et al. (1988) and the diagnostic disc test (DD, version CPD/CD 01) according to Appleton (1999). In the next step examined strains were analysed in two further tests, which are variants of DD method: CAZ/CD 02 test with discs containing ceftazidime and ceftazidime/clavulanic acid, and CTX/CD 03 test with the use of cefotaxime and cefotaxime/clavulanic acid discs. ESBL-positive strains first of all belonged to the species E. coli and K. pneumoniae. In the case of seven analysed strains consistent results of determinations were not obtained with the use of different disc diffusion methods. Application of several disc diffusion methods to determine ESBL-positive strains of gram-negative rods increases the probability of their proper identification.     

Use of new methods for detection of beta-lactamases with extended-spectrum substrates (ESBLs)

The aim of performed examinations was to compare results of two methods applied for the detection of extended-spectrum beta-lactamases (ESBLs). Two hundred and sixty strains of Gram-negative rods were cultured from clinical specimens obtained from hospitalized patients. These strains were identified as ESBL-positive on the basis of the double-disc method (DDST) according to Jarlier et al. (1988) results. Identification of strains was performed in automatic ATB Expression system (bioMerieux, France). All strains were determined using a novel test for detection of ESBL-type enzymes (DD) according to Appleton (1999). Two discs are applied in this test: with cefpodoxime (CPD) and with cefpodoxime/clavulanate (CD 01, diagnostic disc). Consistent results of two methods (DDST and DD) were obtained in the case of 166 from among 260 of examined strains (60.4%). Consistent results concerned 161 out of 222 examined strains of enteric rods (72.5%) and only 5 from among 38 of other strains (mostly belonging to the group of non-fermenting rods). On the basis of performed investigations it can be stated that the novel method of extended-spectrum beta-lactamases (ESBLs) detection (DD) according to Appleton (1999) is more objective and easier for interpretation than the double-disc synergy test (DDST) according to Jarlier et al. (DDST), which is widely applied in the routine microbiological diagnostics.     

In vitro activity of cefepime against ESBL-positive clinical strains of gram-negative rods

The aim of this study was to determine an in vitro activity of cefepime against ESBL-positive clinical strains of Gram-negative rods isolated from hospitalized patients. Experiments were performed with 100 ESBL-positive strains of Gram-negative rods isolated from clinical samples in 2004. Strains were identified with the use of automatic ATB Expression system and biochemical ID 32 GN tests (bioMdrieux sa). Extended-spectrum beta-lactamases (ESBLs) were detected by means of disc diffusion methods: the double-disc synergy test (DDST) and the diagnostic disc test (DD, Oxoid Ltd, UK). Susceptibility in vitro of ESBL producers to 4th generation--cefepime was determined with gradient diffusion method Etest (AB Biodisk, Solna, Sweden). MIC value of cefepime was assessed for each strain. Among 100 ESBL-producing strains, 94--belonged to enteric rods and 6--to nonfermentative rods. The greatest number of strains belonged to the species Serratia marcescens (27% of all strains) and next--to the species Enterobacter cloacae (21%). Fourteen strains were susceptible (S) in vitro to cefepime, 12--intermediately susceptible (I) and 74--resistant (R). Application of cefepime in a therapy of infections caused by ESBL-positive strains of Gram-negative rods highly susceptible in vitro to this antibiotic, should be considered.     

Detection of the expression of extended-spectrum beta-lactamases (ESBL) in clinical strains of Klebsiella pneumoniae and Escherichia coli: comparison of E-test and two-disc methods

The aim of this study was to compare the effectiveness of E-test and two disc methods applied for the detection of extended spectrum beta-lactamases. All strains were tested by E-test, by double-disc synergy test (DDST) according to Jarlier (cefotaxim, ceftazidim, aztreonam and clavulanic acid) and also by disc test according to Appleton (cefpodoxime and cefpodoxime with clavulanic acid, CPD and CD01 disc). We tested 148 clinical strains of E. coli and 78 strains of K. Pneumoniae. In case of K. pneumoniae, the activity of the ESBLs was detected among 30 strains--both in E-test, Jarlier test and Appleton test. Among E. coli, four strains were found ESBL-positive in the test according to Jarlier but only three strain of these when E-test and Appleton test was used. The results of investigations performed suggest, that E-test and disc methods according both Jarlier and Appleton have the same effectiveness in detection ESBLs among K. pneumoniae strains. However, in case of E. coli, interpretation of results may present a problem.     

Antibiotic susceptibility and occurrence of ESBL, IBL and MBL in Pseudomonas aeruginosa strains

The aim of this study was to evaluate the drug susceptibility of P. aeruginosa strains and to detect strains producing inducible beta-lactamases (IBL), extended-spectrum beta-lactamases (ESBL), and metallo-beta-lactamases (MBL). During 6 month (October 2005 - March 2006), 66 strains of P. aeruginosa strains were cultured from clinical specimens obtained from patients of two of hospitals in Siedlce and from patients of outpatient clinics. All the strains were identified in the automatic ATB (bio Mérieux). The susceptibility of bacteria to antibiotics was tested by standard disc diffusion method. The majority of strains were susceptible to meropenem (89.4%), piperacillin combined with tazobactam (84.8%), ciprofloxacin (84.8%) and piperacillin (83.3%). Many of our strains were resistant to carbenicillin (69.7%), mezlocillin (45.5%), gentamicin (42.4%) and netylmicin (30.3%). 6 strains (9.1%) were multidrug-resistant (MDR). Inducible beta-lactamases were detected with the use double disc method according to Sanders and Sanders. ESBL-producing strains were detected with double disc test (DDST) according to Jarlier et al. These strains were identified as ESBL-positive on the basis of the DDST were also determined using a double disc (DD) test according to Appleton. Production of metallo-beta-lactamases (MBL) was examined with the use of Etest MBL (AB Biodisk, Sweden) and the double disc test according to Arakava et al. Sixty-five IBL-producing strains (98.5% of all strains) and three strains (4.5%) with MBL activity were detected. Strains producing extended beta-lactamases (ESBL) were not found.     

High effectiveness of the method with cefpirome in detection of extended-spectrum beta-lactamases in different species of gram-negative bacilli

Gram-negative bacilli were examined for ESBL production by using four methods: double-disc synergy diffusion test (DDST), and three tests of combined discs with cefpodoxime, ceftazidime and cefotaxime alone and the same cephalosporins with clavulanic acid. Strains determined as ESBL-negative with all these tests were examined by using fifth method with cefpirome. 47,5% from 178 negative in other methods strains, appeared ESBL-positive in this test. The examined strains belonged to 16 different species. Most of them were Enterobacter cloaceae, Serratia marcescens, Pseudomonas aeruginosa and Acinetobacter baumanii. It seems that the combined discs method with cefpirome may be usefull for phenotypic detection of ESBL producing bacteria also in the case of strains where ESBL production is camouflaged with derepressed chromosomal AmpC beta-lactamases.     

ESBL-positive strains of the Bacteroides fragilis group isolated from patients at the regional hospital center in Płock (Poland)

The aim of this study was to confirm a presumptive qualification of clinical B. fragilis group strains isolated in P?ock as ESBL-positive strains and to determine some properties of these strains. Twenty four clinical strains belonging to the B. fragilis group, isolated first of all from surgical patients, were received for testing. Identification of strains was performed in the automatic ATB Expression system (bioMerieux sa, France) using biochemical API 20 A strips. Strains were tested for the production of catalase (ID Color Catalase test, bioMerieux sa) and beta-lactamase (Cefinase, BBL, Becton Dickinson, USA). Susceptibility of strains to four antimicrobial agents: clindamycin, metronidazole, amoxicillin/clavulanic acid and imipenem was determined by Etest (AB Biodisk, Sweden). ESBLs were detected with the use of two disc diffusion methods: the double-disc synergy test (DDST) according to Jarlier et al. and the diagnostic disc (DD) test according to Appleton. Seventeen of examined strains belonged to the species Bacteroides fragilis, three--to B. ovatus/thetaiotaomicron, two--to B. distasonis, one--to B. uniformis and one--to B. stercoris/eggerthii. One strain (B. uniformis) did not produce catalase, whereas all strains produced beta-lactamases. Examined strains were susceptible in vitro to metronidazole, amoxicillin/clavulanic acid and imipenem. One clindamycin-resistant strain was detected (B. fragilis). Occurrence of ESBL-type enzymes was confirmed in 22 strains of following species: B. fragilis (17 strains), B. ovatus/thetaiotaomicron (3), B. distasonis (1) and B. uniformis (1). Clinical strains of the B. fragilis group with a new mechanism of resistance to beta-lactam antibiotics appeared during last years in Poland. They produce extended-spectrum beta-lactamases (ESBLs), so they are resistant to penicillins, cephalosporins and monobactams. Monitoring of infections caused by these threatening strains in hospital patients is very important.     

Beta-lactamases with a wide substrate spectrum in gram negative strictly anaerobic rods

This study was performed to determine the susceptibility of the clinical strains of Gram-negative strictly anaerobic rods to newer beta-lactam antibiotics. Also, the trial was undertaken to detect strains producing extended-spectrum beta-lactamases (ESBLs) and inducible beta-lactamases (IBLs) among Bacteroides spp. and Prevotella spp. rods isolated from hospitalized patients. One hundred strains of Gram-negative, obligatory anaerobic rods were applied in the study. The strains were identified in automatic ATB system using API 20 A strips. beta-lactamase-positive strains were determined with disc nitrocefin test. ESBL-producing strains were detected with double disc test according to Jarlier et al. (1988). Clavulanate was applied as the inhibitor of these beta-lactamases (AMO/CLAV disc). ESBL-positive strains were confirmed with the use of E test (TZ/TZL strip). Inducible beta-lactamases were determined by double disc method according to Sanders and Sanders (1979). Cefoxitin was the inducer of these beta-lactamases (FOX disc). Among 93 Bacteroides spp. strains and 7 Prevotella spp. strains, 91 strains (91%) produced beta-lactamases. Two ESBL-producing strains (2%) were detected. Strains producing inducible beta-lactamases (IBL) were not found. A high activity of the examined beta-lactam antibiotics against strains of Gram-negative anaerobes was found. The majority of strains were susceptible to piperacillin (95%), piperacillin combined with tazobactam (99%), ticarcillin combined with clavulanic acid (99%), meropenem (97%) and imipenem (99%). The obtained results indicate the necessity of ESBL determination among strains of the genus Bacteroides, isolated from clinical specimens. Newer beta-lactam antibiotics, especially penicillins in combination with beta-lactamase inhibitors and carbapenems, are useful in empiric therapy of infections caused by Bacteroides spp. and Prevotella spp. anaerobic rods.     

Characteristic of clinical strains of gram-negative obligate anaerobes

The aim of the study was to assess prevalence and antibiotic susceptibility profiles ofGram-negative strictly anaerobic bacteria isolated from clinical specimens taken from hospitalized patients in 2005-2006. Biochemical identification and antibiotic susceptibility were done in an automated system ATB Expression (bioMerieux sa). From 12262 specimens examined 867 strains of obligate anaerobes were isolated. Gram-negative strictly anaerobic bacteria were cultured in number of 138 strains (15,9%). All cultures were performed on Columbia agar and Schaedler agar media (bioMerieux sa) supplemented with 5% sheep blood and incubated at 37 degrees C for 48-120 h in 85% N2, 10% H2, 5% CO2. Most frequently isolated was Bacteroides spp. (41,3%). For this group beta-lactamase activity was evaluated by using nitrocefin disc test (Cefinase BBL, Becton Dickinson and Co., Cockeysville, MD, USA). Production of ESBLs was detected with the use of two disc diffusion methods: the double-disc synergy test (DDST) according to Jarlier et al. and the diagnostic disc (DD) test according to Appleton. ESBLs were produced by 5,3% strains of Bacteroides spp. For all Bacteroides spp. strains MIC values were determined by gradient diffusion method Etest (AB BIODISK, Sweden). ESBLs and MIC were performed on Wilkins-Chalgren solid medium supplemented with 5% sheep blood (Difco Lab., USA) and all plates were incubated at 35 degrees C for 48 hours in 85% N2, 10% H2, 5% CO2. Most Gram-negative obligate anaerobes isolated from clinical specimens are still susceptible to imipenem (100%), metronidazole (99,3%) and beta-lactam antibiotics with beta-lactamase inhibitors: piperacillin/tazobactam (99,3%), ticarcillin/clavulanate (99.3%), amoxicillin/clavulanate (97.8%).     

2010年CLSI头孢菌素折点改变对产ESBLs奇异变形杆菌药物敏感性结果评估和分析

目的评估2009年CLSI M100-S19及2010年CLSI M100-S20文件中头孢他啶（CAZ）、头孢噻肟（CTX）和头孢唑啉（CFZ）最低抑菌浓度新旧折点变化对产ESBLs奇异变形杆菌药物敏感性试验结果的影响。方法对临床分离33株奇异变形杆菌进行产ESBLs菌株的确证试验,琼脂稀释法检测最低抑菌浓度（MIC）,根据药物敏感性结果分别对产ESBLs奇异变形杆菌和非产ESBLs奇异变形杆菌在S19和S20新旧折点下CAZ、CTX和CFZ三种药物的敏感性以及ESBLs阳性菌株分布率进行界定。结果 33株奇异变形杆菌中,产ESBLs菌株6株（18.2%）,由旧折点下耐药率50%（CAZ）、50%（CTX）和66.7%（CFZ）分别上升为新折点下66.7%（CAZ）、100%（CTX）和100%（CFZ）,敏感率由旧折点下33.3%（CAZ）和50%（CTX）分别下降为新折点下16.7%（CAZ）和0%（CTX）,CFZ在新旧折点下均为0,新旧折点标准下药敏结果分布率的差异均有统计学意义。结论对CTX、CFZ,按照CLSI S20新折点琼脂稀释法MICs折点标准判读药敏结果与ESBLs表型检测结果具有高度一致性,临床可以根据药敏结果选择用药,无需进行产ESBLs检测。但对于CAZ,尚需要进一步深入研究和评价。     

Assessment of the susceptibility of the gram-negative rods isolated from hospitalized patients to cefoperazone/sulbactam

The susceptibility to cefoperazone/sulbactam of 197 strains of Gram-negative rods demonstrating an ESBL-positive phenotype was determined. The assortment of the investigated strains was as follows (numbers of strains are given in the brackets): E. cloacae (63), S. marcescens (46), K. pneumoniae (21), P. mirabilis (17), E. coli (9), P. vulgaris (8), P. aeruginosa (20) and A. baumanni (13). 83 strains from 197 were susceptible (42.1%). The MIC values were determined and the disc-diffusion method was performed. The susceptibilities among particular species were as follows (the order of data in the brackets is: % of the susceptible strains/MIC50/MIC90): E. cloacae (54.0/16/64), S. marcescens (23.9/64/> or = 128), K. pneumoniae (38.1/32/64), P. mirabilis (41.2/32/64), E. coli (44.4/32/32), P. vulgaris (75.0/8/32), P. aeruginosa (35.0/32/64), A. baumannii (46.2/32/64). Using disc-diffusion method, for 184 strains the difference between diameter of the inhibition zone around the disc with cefoperazone and the disc with cefoperazone/sulbactam was calculated. This difference amounted 5 mm or more in the case of 76.6% of the investigated strains. The results indicate that the comparison of the inhibition zones around cefoperazone and cefoperazone/sulbactam discs may be an additional method useful for phenotypic detection of ESBL producing organisms. These results highly correlated with results obtained by using analogous test with cefpirome and cefpirome/clavulanic acid (85.6% of concordance).     

临床分离肺炎克雷伯菌耐药性监测

目的了解临床分离肺炎克雷伯菌的耐药性,为临床合理应用抗菌药物提供实验室依据。方法采用微量稀释法对392例临床分离肺炎克雷伯菌进行药物敏感性测定;超广谱β-内酰胺酶(Extendedspectrumbeta-lactamases,ESBLs)检测用微量稀释法初筛,纸片法做确证试验。结果肺炎克雷伯菌对18种抗菌药物的药敏结果中,耐药率大于30%的抗菌药物多达11种;其中氨苄西林-舒巴坦、氨苄西林、头孢噻吩、哌拉西林、复方新诺明和头孢唑啉的耐药率高达20%以上。耐药率低于10%的抗菌药物仅有4种,分别为头孢曲松(7.7%)、头孢噻肟(7.4%)、氨曲南(6.9%)和亚胺培南(3.1%)。其它抗菌药物的耐药率都高于10%。产ESBLs菌株的发生率为32.9%～45.8%,平均为39.8%;产ESBLs菌株对多种抗菌药物的耐药率显著高于非产ESBLs菌株(P<0.05)。结论临床分离肺炎克雷伯菌对多种抗菌药物的耐药率较高,尤其是产ESBLs菌株的高耐药率及多重耐药性更为明显。临床应加强对肺炎克雷伯菌耐药性的监测并预防耐药菌株的传播流行。     

Metallo-beta-lactamase-producing gram-negative rods isolated from inpatients and outpatients, detected using Etest MBL

The aim of presented study was to detect MBL-positive strains in a group of clinical carbapenem-resistant strains isolated from inpatients and outpatients during last four years. From the beginning of November 2001 to the end of October 2005, one hundred and four strains resistant to carbapenem antibiotics--imipenem and meropenem were cultured from clinical samples obtained from patients of the Infant Jesus Clinical Hospital Centre for Trauma Treatment in Warsaw and from patients of outpatient clinics. Strains were identified and their susceptibility to antibacterial agents was determined in the automatic ATB Expression system (bioMérieux). Resistance to imipenem and meropenem was confirmed with a disc diffusion method. Production of metallo-beta-lactamases (MBL) was examined with the use of Etest MBL (AB Biodisk, Sweden), and extended-spectrum beta-lactamases (ESBL) by means of following procedures: DDST and / or DD (four variants) (Oxoid Ltd., England). MBL-positive strains (36) were cultured in cases of infections in adult patients (35 strains) and in a child (1 strain). Majority of strains belonged to the species P. aeruginosa (27), several strains - to the species P. putida (6) and remaining strains--to P. stutzeri, A. xylosoxidans, and E. cloacae (1 strain of each species). Four strains were producers of MBL-type and ESBL-type beta-lactamases. According to our knowledge and accessible literature described strains (except one paediatric strain) are the first MBL-positive strains isolated from adult hospitalized patients and adult ambulatory patients in Poland. Additionally, MBL-positive E. cloacae strain is probably the first MBL producer isolated in Poland, which belongs to the group of enteric rods. MBL-producing strains of Gram-negative rods, detected by phenotypic Etest MBL method, will be verified with genetic procedures.     

产超广谱β- 内酰胺酶肺炎克雷伯菌耐药性及其基因分型分析

目的:了解新疆独山子地区肺炎克雷伯菌超广谱β-内酰胺酶(ESBLs)的发生率、作为指示剂的五种抗生素的检出情况及ESBLs主要基因型。方法:收集临床分离的148株肺炎克雷伯菌,采用双纸片协同筛选法、NCCLS推荐的表型筛选和确证试验对细菌进行ESBLs产酶株的识别;耐药基因的质粒重组、转化,聚合酶链反应(PCR)扩增阳性产物测序,通过GenBank对序确定基因型。结果:本地区肺炎克雷伯菌ESBLs的分离率达31.1%,头孢曲松(CRO)安曲南(ATM)和头孢噻肟(CTX)头孢泊肟(CPD)、头孢他定(CAZ)作为指示剂检出率分别为97.8%、95.6%、93.4%、76.0%、65.2%;本地区产ESBLs菌株耐药基因型CTX-M-22占54.3%,CTX-M-18占41.3%,TEM61.8%,52.1%的产ESBLs肺炎克雷伯菌SHV耐药基因阳性。结论:CRO、ATM和CTX对检测ESBLs阳性率较高;CTX-M-22、CTX-M-18是本地区产ESBLs菌株的主要基因型。     

Antibiotic susceptibility and occurrence of ESBL in Pseudomonas aeruginosa strains isolated from different clinical specimens

The aim of this study was to evaluate the drug susceptibility of 132 P. aeruginosa strains isolated from patients hospitalized in SPSK University Hospital in Bialystok. The isolates were obtained from clinical specimens over an 11-month period in 2001 and 2002. All the strains were identified in automatic ATB system using API 20 NE strips, and their susceptibility to antibiotics was tested by standard disc-diffusion method and agar dilution method. The minimal inhibitory concentration (MIC) was determined for five antibiotics: piperacillin, amikacin, ceftazidime, imipenem and ciprofloxacin. The majority of strains were susceptible to ceftazidime (91.7%), piperacillin combined with tazobactam (85.6%), amikacin (80.3%), meropenem and imipenem (81.8%). Many of our strains were resistant to cefotaxime (73.5%), ticarcillin (53%) and ciprofloxacin (48.5%). Also, the trial was undertaken to detect strains producing extended-spectrum beta-lactamases (ESBL) and inducible beta-lactamases (IBL) among P. aeruginosa rods isolated from different specimens. ESBL-producing strains were detected with double disc test (DDST) and combination double disc (CD) test. Clavulanate was applied as the inhibitor of these beta-lactamases. Strains producing ESBL were not found. On the other hand, as many as 127 P. aeruginosa strains (96.2%) produced inducible beta-lactamases (IBL).     

Conjugative transfer frequency of resistance genes from ESBL-producing Enterobacteriaceae strains isolated from patients hospitalized in pediatric wards

The aim of this study was to evaluate the transfer frequency of plasmids encoding extended-spectrum beta-lactamases (ESBLs) from clinical isolates of Enterobacteriaceae to E. coli K12 C600 recipient strain. Additionally, resistance patterns to antimicrobial drugs of the isolates as well as transconjugants were analyzed. Fifty-four clinical strains belonging to the Enterobacteriaceae family were isolated from children hospitalized in Medical University Hospital in Wroc?aw. All the strains studied were identified in automatic ATB system using ID32E tests. Besides, they were ESBL-positive as was confirmed by the double-disc synergy test (DDST). The minimal inhibitory concentration (MIC) was determined for twelve selected antibiotics and chemotherapeutics. The majority of the strains (87%) were able to transfer plasmid-mediated ESBL to E. coli K12 C600 recipient strain with a frequencies ranged from 10(-5) to 10(-1) per donor cell. All the isolates studied as well as their transconjugants were susceptible to imipenem, meropenem and norfloxacin (MIC <1mg/L). On the other hand, these strains displayed high level of resistance (MIC 512 - >1024 mg/L) to cefotaxime, ceftriaxone, gentamycin, amikacin and cotrimoxazole. Genetic markers conferring resistance to aminoglycosides and cotrimoxazole were often co-transferred to recipient strain in conjugation process.     

CLSI三代头孢菌素新旧折点对判定肺炎克雷伯茵药物敏感性及产ESBLs菌株分布的影响

目的了解CLSI2010（S20）及2009（S19）头孢他啶（CAZ）、头孢噻肟（CTX）新旧折点变化对本地区肺炎克雷伯菌药物敏感性及产超广谱β-内酰胺酶（ESBLs）菌株分布的影响。方法收集安徽医科大学第一附属医院临床分离的50株肺炎克雷伯菌;纸片扩散法测定菌株对CTX及CAZ的敏感性,CLSI表型确证试验确定产ESBLs菌株,改良三维试验检测CTX和／或CAZ耐药、非产ESBLs菌株的产酶情况。结果在S19折点下,CTX和CAZ耐药率分别为54．0％、30．0％;在S20折点下,耐药率分别为68．0％、42．0％。产ESBLs菌株总检出率为64．0％（32／50）。旧折点下,分别有81．3％、43．8％的产ESBLs菌株分布在CTX和CAZ耐药菌株中;新折点下,升高至100％、62．5％。2株对CTX和／CAZ耐药、非产ESBL菌株中,1株同时产ESBLs和AmpC酶,1株仅产AmpC酶。结论根据S20,肺炎克雷伯菌对CTX和CAZ的耐药率较S19均有所增高;并提高了耐药表型与产ESBLs菌株的一致性。产AmpC酶可影响表型确证试验对产ESBLs菌株的检出。     

Occurrence of extended-spectrum β-lactamases among<Emphasis Type="Italic">Escherichia coli</Emphasis> isolates from hospitalized and healthy children

The prevalence of extended-spectrum beta-lactamases (ESBL) was determined among isolates of Escherichia coli (n = 63) isolated from hospitalized (43) and healthy (20) children. Ten isolates (21%) were ESBL-positive for two screening tests, the double disk-synergy test and the Oxoid Combination Disk method. One ESBL-positive isolate came from a healthy child. The transfer frequency of oxyimino-beta-lactam resistance from ESBL-producing isolates to E. coli K12 C600 recipient strain ranged from 10(-8) to 10(-5) per donor cell. Donor strains and transconjugants displayed susceptibility patterns typical of ESBL producers. They were resistant to oxyimino-beta-lactams but susceptible to clavulanic acid and carbapenems. Seven out of the 10 ESBL-positive isolates were found to produce MR/MS fimbria, which may play an important role in the colonization of the human intestinal mucosa.     

同时产多种基因型ESBLs肺炎克雷伯菌的分析

目的分析汕头大学医学院第一附属医院临床分离的肺炎克雷伯菌同时产多种基因型ESBLs的情况。方法采用PCR扩增对产ESBLs的肺炎克雷伯菌初步分型,然后PCR扩增阳性产物克隆测序分析,脉冲场凝胶电泳（PFGE）分型;用浓度梯度法检测10种抗菌药物对同时产多种基因型ESBLs的肺炎克雷伯菌的最低抑菌浓度（MIC）。结果83株产ESBLs的肺炎克雷伯菌中,发现9株同时产TEM、SHV和CTX-M型,测序结果为CTX-M-3、TEM-1及多种SHV型（SHV12及SHV28）。将这9株细菌作PFGE分析,结果共被分成7个型。药敏结果表明,9株菌除对亚胺培南敏感外,对氨曲南、头孢曲松、头孢噻肟、头孢他啶、丁胺卡那和四环素均高度耐药,对哌拉西林／三唑巴坦、头孢吡肟、环丙沙星极少菌株敏感。结论发现CTX—M-3超广谱-及多种SHV型超广谱-和TEM-1广谱β-内酰胺酶基因同时存在该院临床分离的肺炎克雷伯菌中,耐药十分严重,应引起重视。     

革兰阴性杆菌ESBLs和AmpC酶的检测及耐药分析

目的检测引起医院感染的革兰阴性杆菌携带产ESBLs和去阻遏AmpC酶状况,探讨各菌对临床常用抗生素的主要耐药机制及耐药性,为临床制定合理使用抗生素策略提供依据。方法采用全自动微生物分析仪（VITEK-32）做细菌鉴定和药敏试验,用纸片扩散确证法检测超广谱β-内酰胺酶（ESBLs）,用三维法检测高水平表达染色体编码的AmpC酶。结果158株革兰阴性杆菌ESBLs检出率为26．6％,主要菌为大肠埃希菌（45．2％）、肺炎克雷伯菌（42．9％）、阴沟肠杆菌（11．9％）。AmpC酶检出率为10．1％,主要为鲍曼不动杆菌（43．8％）、阴沟肠杆菌（25％）;上述产酶细菌均对青霉素和一、二、三代头孢菌素、磺胺类、喹诺酮类、氨基糖苷类耐药,对亚胺培南敏感。结论革兰阴性杆菌耐药机制主要是产超广谱β-内酰胺酶和AmpC酶,这些产酶菌株均出现多重耐药。