同时产DHA-1型头孢菌素酶和SHV-12型超广谱β内酰胺酶的肺炎克雷伯菌

目的研究头孢菌素酶(AmpC)和超广谱β内酰胺酶(ESBLs)在临床分离肺炎克雷伯菌中的流行、表型及其基因特性。方法先后用标准纸片扩散法、三维试验、等电聚焦、酶抑制试验以及微量稀释法等进行表型检测。然后用接合试验、多重聚合酶链反应(PCR)以及基因测序等方法进行分子生物学研究。结果受试的86株细菌中有4株三维试验阳性，等电聚焦以及酶抑制试验表明这些菌株都产一种等电点(PI)为7．8并具有AmpC性质的β内酰胺酶，基因测序表明和DHA01型AmpC酶一致；它们同时伴随产生一种PI为8．2的ESBLs，基因测序表明其来源为SHV-12。微量稀释法检测表明上述菌株不但对多种三代头孢菌素耐药，而且出现了对头孢吡肟耐药，但对碳青霉烯类抗生素仍敏感。结论本研究发现同时产DHA-1型高产AmpC酶和SHV-12型ESBLs的肺炎克雷伯菌及其耐药表型。此类菌株的出现，将为临床抗感染治疗带来新的困难。     

Prevalence of the genes encoding extended-spectrum beta-lactamases, in Escherichia coli resistant to beta-lactam and non-beta-lactam antibiotics

The prevalences of extended-spectrum beta-lactamases (ESBL) and their encoding bla genes, TEM, SHV and CTX_M, were investigated in isolates of Escherichia coli that were resistant to beta-lactam and/or non-beta-lactam antibiotics. Of the 250 E. coli isolates investigated, all of which came from patients in a major hospital in southern Lebanon, 61 (13.3%) were found to have ESBL, their production of beta-lactamase being confirmed by the ceftazidime and ceftazidime/clavulanic-acid disc methods. All 61 ESBL isolates were resistant to beta-lactams and sensitive to imipenem, piperacillin/tazobactam and cefoxitime. Only 40% were resistant to fluoroquinolones, 33% were resistant to aminoglycosides, and 18% were considered to have multi-drug resistance.The results of the PCR-based amplification of the bla gene in DNA samples from the 61 ESBL isolates indicated that 11 (18%) of the isolates carried both the TEM and SHV genes, 37 (61%) carried the TEM gene but not the SHV, and 13 (21%) had the SHV gene but not the TEM. None of the isolates carried the CTX_M gene. Of the 37 TEM-positive/SHV-negative isolates, 43% were resistant to fluoroquinolones and 37% to aminoglycosides. Increased resistance to non-beta-lactam antibiotics was observed in the isolates harbouring both the TEM and SHV genes, of which 54% were resistant to all of the tested antibiotics except imipenem, 36% were only resistant to fluoroquinolones, and 9.1% only resistant to aminoglycosides.The possibility that the concomitant presence of TEM- and SHV-type beta-lactamases is associated with resistance to non-beta-lactam antibiotics requires further research. The prevalences of ESBL and their encoding genes in Gram-negative bacteria collected from various regions in Lebanon will now be investigated.     

大肠埃希菌和肺炎克雷伯菌中质粒AmpCβ内酰胺酶基因型的检测

目的了解产质粒介导的AmpC酶大肠埃希菌和肺炎克雷伯菌的耐药性和基因型。方法收集2002年1月至2004年5月间呼吸科临床标本中分离的大肠埃希菌和肺炎克雷伯菌共110株，用酶提取物三维试验检测AmpC酶；用等电聚焦电泳、耐药质粒电转化试验、聚合酶链反应（PCR）及测序确定AmpC酶基因型。结果大肠埃希菌和肺炎克雷伯菌中AmpC酶检出率分别为9．30％（4／43），4．48％（3／67）。药敏试验结果显示产酶株对头孢西丁全部耐药，对第三代头孢菌素、酶抑制剂、氨曲南、阿米卡星及环丙沙星均有不同程度耐药，对头孢吡肟及亚胺培南较敏感。7株产AmpC酶菌株中有5株通过电转化试验可将头孢西丁耐药性传递给受体菌，经PCR扩增和测序证实为质粒介导DHA-1型AmpC酶。结论临床分离的大肠埃希菌和肺炎克雷伯菌中已经出现产质粒介导AmpC酶菌株，其耐药性能够水平传播，给临床抗感染治疗带来重大威胁。     

产超广谱β内酰胺酶菌株中质粒头孢菌素酶的研究

目的 调查产超广谱β内酰胺酶(ESBLs)的大肠埃希菌和肺炎克雷伯菌中质粒头孢菌素酶(AmpC酶)的发生率及其基因型。方法 收集北京朝阳医院2001年1～12月头孢西丁耐药的产ESBLs的24株大肠埃希菌、8株肺炎克雷伯菌,采用等电聚焦电泳测定β内酰胺酶的等电点;接合试验证实酶基因有无可转移性;脉冲场凝胶电泳(PFGE)确定耐药株的亲缘关系;对AmpC酶基因进行多重PCR及序列分析确定其基因型。结果 2001年北京朝阳医院ESBLs的发生率,大肠埃希菌为16.8％(49／292),肺炎克雷伯菌为160.5％(35／212);ESBLs株中质粒AmpC酶的发生率,大肠埃希菌为2．0％(1／49),肺炎克雷伯菌为17．1％(6／35)。这7株菌均产生DHA-1型AmpC酶;1株肺炎克雷伯菌可将头孢西丁耐药性传给受菌体。该7株菌都产TEM-1酶、5株产CTX-M-3型ESBL、2株产SHV-12型ESBL;该7株菌携带质粒2～5个,且都有约33～36kb的大质粒。PFGE发现这7株菌来自多个不同的克隆株。结论 北京朝阳医院ESBL阳性的大肠埃希菌和肺炎克雷伯菌中,有7株既产DHA-1型质粒AmpC酶,又产CTX-M-3或SHV-12 ESBL。这7株菌来自多个不同的克隆株。     

Cefotaxime-resistant Citrobacter freundii in isolates from blood in a tertiary teaching hospital in Northern Taiwan

From January 2002 to December 2003, 12 patients in a tertiary teaching hospital in northern Taiwan had bloodstream infections caused by Citrobacter freundii. Seven of the 12 isolates were resistant to cefotaxime. Using polymerase chain reaction and DNA sequencing, 3 of the 7 cefotaxime-resistant C. freundii isolates were found to carry extended-spectrum beta-lactamase (ESBL). AmpC beta-lactamase genes were also detected in all strains of C. freundii. All strains of C. freundii with MICs >or=4 mg/L for cefepime were positive for ESBL. Rather than performing PCR on all cefotaxime-resistant C. freundii isolates, assessment of the MIC for cefepime might be a practical way to choose between treatment with cefepime or with carbapenems.     

Prevalent phenotypes and antibiotic resistance in Escherichia coli and Klebsiella pneumoniae at an Indian tertiary care hospital: plasmid-mediated cefoxitin resistance.

BACKGROUND: The beta-lactam antibiotics, in combination with aminoglycosides, are among the most widely prescribed antibiotics. However, because of extensive and unnecessary use, resistance to these drugs continues to increase. In recent years, resistance in the Indian bacterial population has increased markedly, the majority showing complex mechanisms. Due to increased transcontinental movement of the human population, it would be wise to know the prevalence and resistance complexity of these strains, well in advance, in order to formulate a policy for empirical therapy. METHODS: One hundred and eighty-one isolates of Escherichia coli and 61 isolates of Klebsiella pneumoniae obtained from 2655 non-repeat samples of pus (912) and urine (1743) were studied, and their resistance rates and patterns were noted. The isolates were analyzed for prevalent aminoglycoside and cephalosporin resistance phenotypes and for the presence of extended spectrum beta-lactamase (ESBL) and AmpC enzymes by spot-inoculation and modified three-dimensional tests developed in our laboratory. Fourteen isolates of E. coli and six of K. pneumoniae, resistant to all of the antibiotics tested, were selected for plasmid screening, curing, and transconjugation experiments, and for comparative evaluation of the double disk synergy test (DDST) and modified three-dimensional test (TDT) for detection of beta-lactamases. RESULTS: Urinary E. coli isolates showed maximum susceptibility to amikacin (57.1%), followed by tobramycin (38.5%) and gentamicin (31.9%). Eighteen (19.8%) isolates were susceptible to cefotaxime, whereas 11 (12.1%) were susceptible to ceftriaxone. The K. pneumoniae isolates from urine samples showed maximum susceptibility to tobramycin (63.6%) followed by amikacin (54.5%). Of the K. pneumoniae isolates, 31.8% were susceptible to cefotaxime and 13.6% were susceptible to ceftriaxone. A more or less similar trend of antibiotic susceptibility was noted in E. coli and K. pneumoniae isolates from pus samples. Twenty-six (14.4%) E. coli and 15 (24.6%) K. pneumoniae isolates were found to be ESBL-producers by NCCLS-ESBL phenotypic confirmatory test. Eighteen (9.9%) E. coli and 19 (31.1%) K. pneumoniae isolates were found to be AmpC enzyme-producers by our modified TDT. The simultaneous occurrence of ESBL and AmpC enzymes was noted in 7.7% and 9.8% isolates of E. coli and K. pneumoniae, respectively. CONCLUSIONS: The prevalence of multidrug-resistant bacterial isolates is quite high in our bacterial population. On comparative evaluation of DDST and TDT in resistant isolates, TDT was found to be the better method, detecting ESBLs in 80% of isolates compared to 15% with DDST. A 19.9-kb plasmid was consistently present in all the screened isolates of E. coli and K. pneumoniae, and was inferred to encode cefoxitin and tetracycline resistance based on curing and transconjugation experiments.     

Prevalence of extended-spectrum beta-lactamase and class 1 integron integrase gene intl1 in Escherichia coli from Thai patients and healthy adults

Among 120 Escherichia coli isolates from Thai patients, 37 and 9 isolates were extended-spectrum beta-lactamase (ESBL) and suspected ESBL producers respectively while 5 E. coli isolates from 120 Thai healthy adults were suspected ESBL producers. Integrase (intl1) gene was detected in 99% of the clinical and 87% of the non-clinical isolates. Among 37 ESBL producers, percent recovery of bla(TEM), bla(CTX-M), bla(SHV) and bla(VEB) was 78%, 78%, 8% and 8%, respectively. Twenty-five isolates of ESBL producers carried both bla(TEM) and bla(CTX-M), 2 isolates carried 3 genes (bla(TEM), blac(CTX-M), and bla(SHV)) and 3 showed no detectable bla gene. Among the 14 suspected ESBL producers, intl1 and bla(TEM) were detected in 13 isolates. ESBL producers from clinical samples were resistant to most of the tested antimicrobial agents compared to non-ESBL producers and isolates from healthy adults with about half of the latter susceptible to all tested antimicrobial agents. Only one clinical isolate was resistant to imipenem. Susceptibility to trimethoprim/sulfamethoxazole among the clinical isolates in ESBL producer group (27%) and non-producer group (33%) were comparable, whereas the percent susceptibility of the non-clinical isolates was about twice that of the clinical isolates.     

CTX-M-3 type beta-lactamase producing Shigella sonnei isolates from pediatric bacillary dysentery cases

In this study, 153 clinical isolates of Shigella were screened for extended-spectrum beta-lactamase (ESBL) production by double-disk synergy test. After being confirmed by the combined disk-test and inhibitor-combined E-test strips, all positive isolates were tested for the bla gene by PCR and the enzymes by isoelectric focusing. DNA sequencing of PCR products revealed that five Shigella sonnei isolates had CTX-M-3 type ESBL enzymes. All of them were resistant to ampicillin, sulfamethoxazole and cefotaxime but susceptible to ofloxacin and ceftazidime. All isolates displayed identical patterns in pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR. This study reports multidrug-resistant (MDR) S. sonnei isolates producing CTX-M-3 type ESBL from successive pediatric bacillary dysentery patients, indicating widespread and rapid spread of CTX-M type ESBL in Shigella spp. To counter this emerging threat to public health, the surveillance of CTX-M type beta-lactamases should be considered, together with measures designed to prevent outbreaks of MDR Shigella in the community.     

Mutations of gyrA gene and parC gene in fluoroquinolone-resistant Escherichia coli isolates from sporadic diarrheal cases

We studied 107 isolates of Escherichia coli O153 from sporadic diarrhea cases in Fukui, Toyama, Aichi, and Saga prefectures from 1991 to 2005 for antimicrobial susceptibility and mechanisms of fluoroquinolone resistance, based on standard disk diffusion. Of 12 drugs tested, ampicillin displayed resistance to 72.9% of isolates, streptomycin to 48.6%, tetracycline to 46.7%, sulfisoxazole to 46.7%, trimethoprim/sulfamethoxazole to 29.9%, nalidixic acid (NA) to 29.9%, and ciprofloxacin (CPFX) to 24.3%. Ten of 32 isolates resistant to 3-6 drugs and 16 of 18 isolates resistant to 7-10 drugs were resistant both to NA and CPFX. Mutations of amino acid in quinolone resistance-determining regions of gyrA and parC genes were detected in 24 isolates resistant both to NA and CPFX, and in 1 isolate resistant to NA. The former possessed a combination of double substitution (S83L and D87L) in GyrA and a single substitution (S80I) in ParC. Some 12 of 24 isolates possessed another single substitution (E84V or E84G or A108T) in ParC. The 25 isolates were classified into 4 types as follows. 1 isolate as type 1: GyrA (S83L) and ParC (S80I); 12 isolates as type 2: GyrA (S83L and D87N) and ParC (S80I); 8 isolates as type 3: GyrA (S83L and D87N) and ParC (S80I and E84G/S80R and E84V); and 4 isolate as type 4: GyrA (S83L and D87N) and ParC (S80I and A108T). In the relationship between amino acid mutations and minimal inhibitory concentrations (MIC) of fluoroquinolone, MICs of CPFX, ofloxacin, and norfloxacin showed 1microg/mL, 2microg/mL and 8microg/mL in type 1; 8 approximately 32microg/mL, 8 approximately 32microg/mL and 16 approximately 256microg/mL in type 2; and 32 approximately 256microg/mL' 32 approximately 128microg/mL and 128-->512microg/ mL in types 3 and 4. These results suggest that most of multiple-antimicrobial-resitant E. coli O153 isolates from sporadic diarrhea cases were resistant to fluoroquinolones and possessed mutations at gyrA and parC genes associated with fluoroquinolone resistance.     

ESBL genotypes in fluoroquinolone-resistant and fluoroquinolone-susceptible ESBL-producing Escherichia coli urinary isolates in Manitoba

OBJECTIVE: Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli are increasingly common in nosocomial and community settings. Furthermore, fluoroquinolone (FQ) and even multidrug resistance (MDR) appear to be associated with certain ESBL genotypes. The purpose of the present study was to determine which ESBL genotypes are associated with FQ and MDR in E coli urinary isolates in Manitoba. METHODS: The authors determined the antimicrobial susceptibility, genetic similarity and ESBL genotype of 27 FQ-resistant and seven FQ-susceptible, ESBL-producing urinary isolates submitted to the clinical microbiology laboratories of two teaching hospitals between October 2000 and April 2005. Susceptibilities to beta-lactams, FQs, trimethoprim-sulfamethoxazole (SXT), doxycycline (DOX), gentamicin (GM) and tigecycline were determined by microbroth dilution; pulsed-field gel electrophoresis (PFGE) was used to determine genetic relatedness, and ESBL genotype was determined by polymerase chain reaction and sequencing. RESULTS: Of 34 ESBL-producing organisms, 27 (79.4%) were found to be ciprofloxacin (CIP) resistant, 27 (79.4%) were SXT resistant, eight (23.5%) were GM resistant and 29 (85.3%) were DOX resistant. Twenty-three (67.6%) had MDR, with concomitant resistance to CIP and SXT; 16 had concomitant resistance to CIP, SXT and DOX; and seven (20.6%) had MDR, with concomitant resistance to CIP, SXT, DOX and GM. All isolates were susceptible to tigecycline. Of 27 FQ-resistant ESBL-producing organisms, seven (25.9%) were genotype CTX-M-14, 19 (70.4%) were genotype CTX-M-15 and one (3.7%) was genotype CTX-M-24. Among the seven FQ-susceptible strains, three (42.8%) expressed SHV-type enzymes, three (42.8%) expressed TEM-type enzymes and one (14.3%) expressed CTX-M-9. CTX-M-15 was the most common MDR-associated genotype. Of a total of 19 strains, 18 (94.7%) were resistant to FQs and SXT; 15 (78.9%) were resistant to FQs, SXT and DOX; and five (26.3%) were resistant to FQs, SXT, DOX and GM. PFGE analysis revealed genetic similarity within CTX-M-15-producing isolates only. CONCLUSION: CTX-M-15 in E coli is strongly associated with an MDR phenotype compared with other genotypes. CTX-M-14 is associated with FQ resistance only. PFGE suggests clonality of CTX-M-15-producing isolates within and among hospitals.     

Molecular characteristics of extended-spectrum β-lactamases in clinical isolates of Enterobacteriaceae from the Philippines

β-Lactamases, including extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases, are major resistance mechanisms of Enterobacteriaceae. Emergence of plasmid-mediated quinolone resistance (PMQR) determinants in ESBL-producing isolates poses a global threat. The molecular characterisitcs of ESBL and PMQR determinants in the Philippines are not well characterized. In this study, we investigated ESBLs and AmpC β-lactamases in clinical isolates of Enterobacteriaceae from the Philippines, and analyzed the association between ESBL and PMQR genes. A total of 91 amoxicilin non-susceptible Enterobacteriaceae were collected at the Research Institute for Tropical Medicine of the Philippines from 2006 to 2008. AmpC- or ESBL-producing isolates were screened by detecting a zone diameter for cefoxitin ≤ 14 mm or cefpodoxime ≤ 20 mm, respectively. Possible ESBL-producing strains were assessed by the ESBL confirmation test of the Clinical and Laboratory Standards Institute. PCR and sequencing were performed to detect the ESBL and PMQR genes. The number of ESBL-producers and AmpC-producers confirmed phenotypically was 17 (18.7%) and 61 (67.0%), respectively. Of 17 phenotypic ESBL-producers, 14 isolates had ESBL genes, including 6 of Escherichia coli, 3 of Enterobacter cloacae, 2 of Enterobacter aerogenes, 2 of Klebsiella pneumoniae, and 1 of Klebsiella ozaenae. Among these isolates, there were 13, 4, and 12 with bla(CTX-M), bla(SHV), and bla(TEM), respectively. Of the bla(CTX-M)-positive isolates, bla(CTX-M-15) shows the highest prevalence, followed by bla(CTX-M-3) and bla(CTX-M-14). Of 14 ESBL-producers identified by PCR, 4, 6, and 7 isolates were positive for qnrB, qnrS, and aac(6')-Ib-cr, respectively. The frequency of aac(6')-Ib-cr positivity was significantly higher among CTX-M-15-producing isolates. Thus, we identified bla(CTX-M), aac(6')-Ib-cr, and qnr in ESBL-producing Enterobacteriaceae from the Philippines, and revealed a significant association between bla(CTX-M-15) and aac(6')-Ib-cr. Local epidemiological data are important for implementing appropriate antimicrobial therapy and effective infection control measures. Continuous monitoring of antimicrobial resistance genes in the Philippines will be required.     

Detection and molecular genetics of extended-spectrum beta-lactamases among cefuroxime-resistant Escherichia coli and Klebsiella spp. isolates from Finland, 2002-2004

Extended-spectrum beta-lactamase (ESBL) producing Escherichia coli and Klebsiella spp. isolates are spreading and becoming an increasing problem concerning treatment, diagnostics and hospital hygiene. We wanted to discover which genotypes are occurring in Finland and to assess the CLSI screening method. The isolates were collected from 26 laboratories during a 3-y period from 2002 to 2004. We studied the zone diameters by disk diffusion according to CLSI recommendations. ESBL genes were detected by PCR and the TEM and SHV genes were sequenced traditionally, while the CTX-M isolates were analysed with pyrosequencing. Of the 402 isolates included in the study, 269 (67%) were confirmed to be ESBL producers according to the CLSI criteria. The CTX-M genes were the most prevalent, especially the combination of a CTX-M-1-group and a TEM-1 gene. In our material there were few isolates that had an ESBL gene but were negative in the CLSI ESBL confirmatory test. During recent y especially the CTX-M producing isolates have increased in Europe and now they are also found in Finland with increasing prevalence.     

Epidemiology of extended-spectrum beta-lactamase producing gram-negative bacilli at Siriraj Hospital, Thailand, 2003

A cross-sectional study was conducted from August to September, 2003 to determine the prevalence and risk factors in acquiring extended-spectrum beta-lactamase (ESBL) producing gram-negative bacilli (GNB) in patients admitted to Siriraj Hospital and the outcomes of these infections. Of 346 isolates of gram-negative bacteria in 249 patients, 102 isolates from 87 patients were colonization only, but 244 isolates from 162 patients were infections. The common GNB were Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii and Enterobacter cloacae. The overall prevalence of ESBL producers was 30.1%. K. pneumoniae had a very high prevalence of ESBL producers (56.9%). The urinary tract was the most common site for ESBL- producing GNB infections. Nosocomial infections, duration from admission to infection, peripheral line, urinary catheterization, nasogastric tube insertion and previous use of beta-lactams, cephalosporins and fluoroquinolones were associated with acquiring ESBL-producing GNB infections. ESBL-producing GNB were significantly more resistant to antimicrobial agents. More than 80% of ESBL-producing GNB were susceptible to carbapenems. Mortality in patients infected with ESBL-producing GNB (41.3%) was significantly higher than those infected with non- ESBL-producing GNB (19.8%).     

Multidrug-resistant human and animal Salmonella typhimurium isolates in France belong predominantly to a DT104 clone with the chromosome- and integron-encoded beta-lactamase PSE-1

Epidemiologic relationships were investigated in 187 ampicillin-resistant Salmonella typhimurium strains (86 human, 101 animal) from >2000 strains isolated in 1994. Of 23 resistance patterns, the most frequent (ampicillin [Am], chloramphenicol [Cm], tetracycline [Tc], streptomycin and spectinomycin [Sm], and sulfonamides [Su]) was found in 69.5% of human and 64.8% of animal isolates. Four beta-lactamase genes were identified, blaTEM (24%), blaPSE-1 (78%), and blaSHV and oxa-2 (each <3%). blaPSE-1 and the integrase gene, intI1, but not blaTEM, blaSHV or oxa-2, were chromosomeborne and found almost exclusively in the AmCmTcSmSu strains. In these, polymerase chain reaction mapping revealed two distinct integrons carrying blaPSE-1 or aadA2. Lysotypes, plasmid profiles, and restriction fragment length polymorphisms (IS200) were determined for 50 representative isolates and for 3 DT104 strains from the United Kingdom (UK). The phage type of the PSE-1-producing AmCmTcSmSu strains was 12 atypic, indistinguishable from that of the DT104 strains. The combined data indicate that the same multiresistant clone has spread through human and animal ecosystems in the UK and France.     

Multidrug-resistant and extended-spectrum beta-lactamase (ESBL)-producing Salmonella enterica (serotypes Typhi and Paratyphi A) from blood isolates in Nepal: surveillance of resistance and a search for newer alternatives.

OBJECTIVES: We evaluated the prevalence of multidrug resistance (MDR) and production of extended spectrum beta-lactamase (ESBL) by Salmonella enterica (serotypes Typhi and Paratyphi A) in a teaching hospital in Nepal. The MDR strains of S. enterica were also tested for susceptibility to newer antibiotics. METHODS: Blood cultures were obtained from 4105 patients with febrile illnesses. Isolates of S. enterica were serotyped and antibiotic susceptibility testing was carried out using disk diffusion (Kirby-Bauer) and E-tests. ESBL screening and phenotype confirmation were done following National Committee for Clinical Laboratory Standards (NCCLS) recommendations for Escherichia coli. RESULTS: A total of 541 isolates of S. enterica serotypes Typhi (47%) and Paratyphi A (53%) were grown. Twenty-eight isolates (5%) of S. enterica were resistant to two or more antibiotics (MDR isolates), with a greater prevalence among serotype Paratyphi A (7%). All ESBL producers (three isolates) were serotype Paratyphi A. Most of the MDR S. enterica showed reduced susceptibility to ampicillin, chloramphenicol, trimethoprim-sulfamethoxazole, ofloxacin, and ciprofloxacin, and had good susceptibility to extended-spectrum cephalosporins and carbapenems. Among the fluoroquinolones, gatifloxacin demonstrated better in vitro activity compared to levofloxacin, ciprofloxacin, and ofloxacin. CONCLUSIONS: A greater prevalence of S. enterica serotype Paratyphi A with higher rates of multidrug resistance and ESBL production is concerning for natives as well as travelers in Nepal since the current typhoid vaccines do not provide protection against this serotype.     

Drug sensitivity test of Campylobacter jejuni strains isolated from human diarrheal stools, chicken meat, and chicken feces, and gene mutation of quinolone-resistant strains

In basic studies on campylobacteriosis, we tested 53 strains from human diarrhea stools and 102 strains from chicken meat and feces obtained between 2002 and 2006 for drug sensitivity to different drugs and gene mutation in quinolone-resistant strains. 1) Of 15 drugs tested, all were resistant to one or more of the following 10 drugs: CEX, 99.4%: ABPC, 59.4%; NA, 40.6%; NFLX, 40.0%; TC and CPFX, 39.4%; PIPC, 38.1%; MINO, 30.3%; KM, 3.2%; and SM, 2.6%. 2) Of 155 drug-resistant strains, 28 (18.1%) were resistant to single drugs and 127 (81.9%) were resistant to multiple drugs. The most frequent pattern of multipledrug resistance was ABPC/PIPC/CEX, followed by ABPC/PIPC/CEX/TC/MINO/NA/NFLX/CPFX. 3) Mutation of GyrA (Thr86 --> Ile) was detected in 43 (97.7%) of 44 quinolone-resistant strains. We found that resistance to beta-lactams, quinolones, and tetracycline antibiotics was high, and most resistant strains were resistant to multiple drugs. We also found that most quinolone-resistant strains had GyrA mutation.     

耐头孢西丁革兰阴性杆菌高产AmpC酶发生率及其基因型和耐药性

目的了解耐头孢西丁革兰阴性杆菌高产AmpC酶发生率及其基因型分布和耐药性状况。方法收集2004年9月-2005年3月分离自南京军区福州总院、福建省立医院、解放军476医院、福州?第二医院住院患者155株耐头孢西丁无重复革兰阴性杆菌,采用酶提取物三维试验检测AmpC酶;API药敏试验板和K-B法测定高产AmpC酶菌株对抗菌药物的敏感性;质粒转化试验定位耐药基因;PCR通用引物扩增AmpC酶与ESBLs基因及其序列测定,确定其基因亚型。结果在155株菌中有36株高产AmpC酶,高产AmpC酶发生率23.2%,分布在13种菌种中,其中大肠埃希菌、肺炎克雷伯菌、鲍氏不动杆菌和阴沟肠杆菌的发生率,分别为29.3%、13.9%、6/10、2/6。在36株高产AmpC酶菌株中检测出DHA-1型2株(肺炎克雷伯菌)、CMY-2型4株(大肠埃希菌),新发现基因CMY-22型1株(大肠埃希菌,GenBank登录号:DQ256079),5株携带CMY基因的大肠埃希菌分别同时带有TEM-1型广谱酶,TEM-144(新发现基因,GenBank登录号:DQ256080)、CTX-M-27、和CTX-M-14超广谱β-内酰胺酶。高产AmpC酶菌株耐药性严重,且呈多重耐药,但对亚胺培南和美罗培南的敏感率>87%。结论耐头孢西丁革兰阴性杆菌高产AmpC酶发生率较高,菌种分布较宽,耐药性强,治疗相关菌造成的感染应以亚胺培南和美罗培南为首选药物。福州地区临床分离株发现产DHA-1型AmpC酶肺炎克雷伯菌、产CMY-2型和CMY-22型AmpC酶大肠埃希菌。CMY-2型和CTX-M-27型为中国大陆首次报告,CMY-22型和TEM-144型为国内外首次发现。     

Prevalence of class A and AmpC β-lactamases in clinical Escherichia coli isolates from Pakistan Institute of Medical Science, Islamabad, Pakistan

In this study, 121 Escherichia coli samples isolated from clinical specimens obtained from Pakistan Institute of Medical Science, Islamabad, Pakistan, were analyzed for extended-spectrum β-lactamases (ESBLs) and AmpC β-lactamases using disk-diffusion assay and polymerase chain reaction. Of the isolates, 78 and 43 were identified as ESBL and AmpC producers, respectively. The highest resistance (89%) was observed against cefotaxime, followed by ciprofloxacin (87.6%) and cefepime (87%). Genetic analysis showed the presence of different class A and class C β-lactamase genes, either alone (44.7%) or in combination (53.6%). CTX-M (57.7%) was the most prevalent among class A, followed by TEM (20.3%) and SHV (15.4%). CIT (including LAT-1 to LAT-4, CMY-2 to CMY-7, and BIL-1) and MOX (including MOX-1, MOX-2, CMY-1, and CMY-8 to CMY-11) family-specific plasmid-mediated AmpC β-lactamases were the most prevalent among these isolates. Our study showed that both class A and class C β-lactamases contributed to cephalosporin resistance in the E. coli isolates, thereby limiting therapeutic options. Co-expression of these enzymes may further hinder the identification of ESBLs, which is a critical step for designing a successful treatment for multidrug-resistant E. coli.     

Epidemiology of ciprofloxacin resistance and its relationship to extended-spectrum beta-lactamase production in Klebsiella pneumoniae isolates causing bacteremia.

A prospective study of Klebsiella pneumoniae bacteremia was performed in 12 hospitals in 7 countries. Of 452 episodes of bacteremia, 25 (5.5%) were caused by K. pneumoniae that was resistant in vitro to ciprofloxacin. Extended-spectrum beta-lactamase (ESBL) production was detected in 15 (60%) of 25 ciprofloxacin-resistant isolates, compared with 68 (16%) of 427 ciprofloxacin-susceptible strains (P=.0001). Multivariate analysis revealed that risk factors for ciprofloxacin resistance in K. pneumoniae included prior receipt of a quinolone (P=.0065) and an ESBL-producing strain (P=.012). In all, 18% of ESBL-producing isolates were also ciprofloxacin-resistant. Pulsed-field gel electrophoresis showed that 11 of the 15 ciprofloxacin-resistant ESBL-producing strains belonged to just 4 genotypes, suggesting that patient-to-patient transmission of such strains occurred. The close relationship between ESBL production and ciprofloxacin resistance is particularly worrisome because the first reported instance of plasmid-mediated ciprofloxacin resistance has been in an isolate of K. pneumoniae also possessing an ESBL.     

Prevalence of multiresistant Gram-negative organisms in a surgical hospital in Ho Chi Minh City, Vietnam

OBJECTIVE: To determine resistance patterns of multiresistant Gram-negative organisms at a surgical hospital in Ho Chi Minh City, Vietnam, in order to guide appropriate antibiotic prescribing and improve infection control procedures. METHOD: All samples sent in for microbiological analysis over a 3-month period were included. A resource neutral double disc-diffusion test was introduced to detect the presence of extended-spectrum beta-lactamase (ESBL) production. RESULTS: We obtained 350 bacterial isolates from clinical specimens; 87.4% were Gram-negative bacteria (GNB). Of these, 88.9% were Enterobacteriaceae, of which 14.7% produced ESBL. Fifteen (37.5%) of these were isolated within 48 h of admission. Resistance to gentamicin and ciprofloxacin occurred in 70.0% and 72.5% of those organisms that produced ESBL and in 39.5% and 38.7% of those that did not. Resistance to third-generation cephalosporins was common: 36.7% of all GNB were resistant to ceftriaxone, 34.0% to cefotaxime, 19.6% to ceftazidime and 36.7% to cefoperazone. CONCLUSION: Multiresistant Gram-negative organisms are common and pose a challenge to antibiotic therapy. Successful implementation of a simple test to detect ESBL production allowed reporting of these organisms, appropriate antibiotic prescribing and infection control interventions. Development of antibiotic-prescribing guidelines must take into account these resistance patterns.