1998～2000年954株烧伤感染菌耐药性调查分析

目的 调查烧伤创面分离菌对抗菌药物的耐药状况。方法 收集临床分离的致病菌，采用纸片扩散法进行药敏试验。结果 在检出的954株细菌中，金黄色葡萄球菌居首位，检出率为27．5％；其次为铜绿假单胞菌，检出率为19．5％；以后依次为表皮葡萄球菌、粪肠球菌、肺炎克雷白杆菌和阴沟肠杆菌，检出率分别为10．4％、6．8％、4．1％和2．6％：万古霉素对金黄色葡萄球菌100％敏感；铜绿假单胞菌对第三代头脑菌素和亚胺培南耐药性显著增加；除铜绿假单胞菌外，其它细菌对阿米卡星的耐药率都有不同程度的降低。结论 细菌耐药性日益严重，随时进行细菌耐药性监测、合理应用抗生素是控制和延缓细菌耐药性的关键。     

Phenotypes of isolates of Pseudomonas aeruginosa in a diabetes care center

BACKGROUND: Pseudomonas aeruginosa is an invasive organism that frequently causes severe tissue damage in diabetic foot ulcers. A major problem in P. aeruginosa infection may be that this pathogen exhibits a high degree of resistance to a broad spectrum of antibiotics. Some researchers feel that P. aeruginosa is a homogeneous species, whereas others have suggested that they are panmictic. Here we characterized P. aeruginosa populations isolated from diabetic foot ulcer and from hospital environment specimens, both from a tertiary diabetes care center in Chennai, India. METHODS: Phenotypic methods like antibiotic susceptibility determinations using Kirby-Bauer's disc diffusion test and minimum inhibitory concentration (MIC) as well as outer membrane protein SDS-PAGE analysis of P. aeruginosa were performed. RESULTS: Twenty three isolates (29.8%) of P. aeruginosa from 77 diabetic foot ulcers and two environmental isolates (13.3%) from 15 different hospital fomites were detected. Both environmental isolates were sensitive to antibiotics than those isolated from clinical specimens by Kirby-Bauer's disk-diffusion method, which correlated the resistance levels by MIC determination. Outer membrane proteins (OMP) corresponding to 21, 23, 43, 46, 50, and 70 kDa were detected. CONCLUSIONS: The study is captivative as the resistance in P. aeruginosa from diabetic foot ulcers seems very common and because all the isolates were resistant to at least one or more antibiotics tested. Disk-diffusion and MIC results shows that piperacillin, amikacin and imipenem retain high levels of antipseudomonal activities and amikacin two times more active than the aforementioned antibiotics to enable itself as a potent antipseudomonal agent in diabetic foot infections. The OMP profile has revealed that clinical isolates were different from hospital environment isolates, which suggests that the origin of infections by P. aeruginosa is mainly due to growth of bacterial strains acquired by patients prior to hospital admission.     

烧伤创面绿脓假单胞菌的分离及对哌拉西林/他唑巴坦耐药情况分析

目的：探讨烧伤创面绿脓假单胞菌对哌拉西林/他唑巴坦（PIP／TAZ)的耐药性并与其他几种治疗绿脓假单胞菌感染的抗生素比较，寻找最佳的治疗用药。方法：对烧伤创面分离出的189株绿脓假单胞菌，用MI-CROSCAN AS4细菌鉴定及药敏分析系统进行鉴定，用K—B法进行药物敏感试验。结果：几种常用于治疗绿脓假单胞菌的药物中，敏感性较高的依次为PIP／TAZ、哌拉西林、头孢他啶、氨曲南和泰能，其耐药率分别为27．5％、42．9％、52．3％、57．1％、74．6％。结论：PIP／TAZ为治疗烧伤创面绿脓假单胞菌较理想的抗生素之一。     

Epidemiologic Study of Pseudomonas aeruginosa in critical patients and reservoirs

BACKGROUND: Pseudomonas aeruginosa is a common cause of nosocomial infections, particularly in intensive care units (ICUs). The aim of this study was to characterize P. aeruginosa clinical isolates by comparing antimicrobial susceptibility patterns with the presence of plasmids and to establish the clonal relatedness by pulsed-field gel electrophoresis (PFGE) typing. METHODS: The patients included those with isolation of P. aeruginosa hospitalized for more than 48 h in the ICU from April to May 1998. Environmental and staff cultures were obtained simultaneously. Minimal inhibitory concentrations, plasmid DNA profiles, and PFGE genomic patterns of enzyme restriction chromosomal DNA were compared. RESULTS: Sixty P. aeruginosa isolates were obtained from 197 clinical specimens, 178 environmental samples, and 47 hand cultures of personnel. Antimicrobial resistance was as follows: tobramycin 100%; ticarcillin, cefotaxime, ceftriaxone, ceftazidime, and gentamicin 80%; cefepime 60%; amikacin, ticarcillin/clavulanate, imipenem, and meropenem 40%; piperacillin and norfloxacin 20%; carbenicillin 12%, and ciprofloxacin 0%. Plasmids were detected in 11 isolates (18%). PFGE typing showed that 23 isolates belonged to a common clone (pattern A), identified from five patients, two nurses, and 10 environmental samples. Ten isolates were grouped in four clusters and 27 isolates had unrelated genomic patterns. There was no relationship among DNA genomic patterns, plasmid profiles, and susceptibility patterns. CONCLUSIONS: PFGE demonstrated the existence of a common clone in a critical care area. Reinforcement of infection control measures is needed to avoid horizontal transmission and severe infections.     

某综合医院铜绿假单胞菌2005-2009年耐药性的变迁

目的了解某综合医院2005-2009年铜绿假单胞菌临床分离株的耐药性变迁,为合理应用抗菌药物提供依据。方法应用回顾性调查方法,对广东省汕头市某综合医院2005-2009年住院病人送检标本中分离的铜绿假单胞菌的药敏试验结果进行统计分析。结果 5年间共检出铜绿假单胞菌1809株,总的分离率为16.34%;标本分布以痰液来源为主,占84.9%;几年间铜绿假单胞菌对氨苄西林、复方新诺明、呋喃妥因、头孢唑啉的耐药率变化不大,对阿米卡星、庆大霉素、妥布霉素、亚胺培南、头孢他啶、头孢吡肟、环丙沙星、左旋氧氟沙星等抗菌药物的耐药率均在2007年达到高峰,在2008、2009年两年间呈连续下降趋势。从2009年的情况来看,耐药率最低的是妥布霉素,为28.97%(168/580),其次为阿米卡星和左旋氧氟沙星,分别为32.07%(186/580)和33.97%(197/580)。结论铜绿假单胞菌对大多数抗菌药物的耐药率有所下降,但总体耐药情况还是比较严重。     

Relationship between clinical and environmental isolates of Pseudomonas aeruginosa in a hospital setting

BACKGROUND: Populations of Pseudomonas aeruginosa have been extensively studied, although there is no general agreement concerning their genetic structure. It has been proposed that P. aeruginosa is a very homogeneous species with 90% of individuals within the same clonal group; nonetheless, other results suggested that Pseudomonas populations are panmictic. Here we compared P. aeruginosa populations from clinical and environmental samples, both isolated from the Bellvitge Hospital of the University of Barcelona in Spain. METHODS: Antibiotic susceptibility determination as well as whole cell and outer membrane protein denaturing gel electrophoresis, pulsed-field electrophoresis, and random amplified polymorphic DNA analysis were performed. RESULTS: Environmental isolates were much more susceptible to antibiotics than those isolated from clinical specimens. The remainder of the analyses revealed high degree of diversity. CONCLUSIONS: Whole-cell proteins, outer-membrane proteins, and pulsed field electrophoresis did not support a close relationship between clinical and environmental isolates. Random amplified polymorphic DNA (RAPD) confirmed the distance between isolates from both sources. This suggests that the origin of hospital infections by P. aeruginosa is due mainly to growth of bacterial strains acquired by patients prior to hospital admission or from patient-to-patient through healthcare workers (HCWs).     

Identification and Distribution of the Clinical Isolates of Imipenem-resistant Pseudomonas aeruginosa Carrying Metallo-β-lactamase and/or Class 1 Integron Genes

To investigate the distribution of the genes of two major metallo-β-lactamases (MBL; i.e., IMP and VIM) and class 1 integrons (intI) in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for blaIMP-1, blaVIM and blaVIM-2 genes. The MBL-positive isolates were further assessed for class 1 integrons by PCR using specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the blaVIM gene was found in 81.5% (53/65) of all isolates, bla gene was found in only 1 isolate and the intI gene was observed in 45.3% (24/53) of blaVIM-positive isolates. One isolate carried simultaneously both blaIMP-1 and intI genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM β-lactamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P. aeruginosa.     

Dominant cagA/vacA genotypes and coinfection frequency of H. pylori in peptic ulcer or chronic gastritis patients in Zhejiang Province and correlations among different genotypes, coinfection and severity of the diseases

Background Almost half of the world’s population suffer from the Helicobacter pylori (H. pylori) infection, but only some individuals develop gastric diseases with clinical symptoms. One reason for the phenomenon may be the different pathogenicity of infected H. pylori strains. The presence of cytotoxin-associated gene A (cagA) and expression of vacuolating cytotoxin activity encoded by vacuolating cytotoxin gene A (vacA) are considered the two major virulent markers of H. pylori. The aim of this study was to detect dominant cagA/vacA genotypes and coinfection frequency of H. pylori in patients with peptic ulceration (PU) or chronic gastritis (CG), and to determine correlations among different cagA/vacA genotypes, coinfection and severity of the diseases. Methods For each of 139 patients in Zhejiang Province who had been diagnosed as PU or CG based on clinical symptoms and gastroscopy, two gastric biopsy specimens (one from antrum and the other from corpus) for H. pylori isolation were taken by two different disinfected biopsy forceps. One hundred and fifty-six H. pylori strains were isolated from both the antrum and corpus biopsy specimens of 78 patients (36 PU and 42 CG). PCRs were performed to detect cagA genes, and signal (s) and middle (m) regions of vacA genes in the H. pylori isolates. The amplified fragments of dominant vacA gene s and m subtypes from representative H. pylori isolates were sequenced after TA cloning. Dominant cagA/vacA genotypes of the H. pylori isolates, coinfection frequency and correlations among the different genotypes, coinfection and severity of the diseases were determined.Results Of the H. pylori strains isolated from the antrum specimens, 96.2% were cagA gene positive, as were 97.4% of the H. pylori strains isolated from the corpus specimens. Only one s region subtype (s1a) and four m region subtypes m1, m2, m1b and m1b-m2 of vacA gene were found. The proportions of vacA gene subtypes s1a/m1, s1a/m2, s1a/m1b and s1a/m1b-m2 in the 83 strains isolated from the antrum specimens were 7.2%, 61.5%, 30.1% and 1.2%, respectively, while those in the other 84 strains isolated from the corpus specimens were 9.5%, 58.3%, 28.6% and 3.6%, respectively. s1a/m2 (58.3% vs 30.1%, χ2=13.47, P<0.01) and then s1a/m1b (28.6% vs 9.5 %, χ2=9.88, P<0.01) were the dominant vacA gene subtypes in the H. pylori isolates. The dominant H. pylori genotype was cagA+s1a/m2 (59.0% from antrum specimens and 57.1% from corpus specimens), and followed by cagA+s1a/m1b (28.9% from antrum specimens and 27.4% from corpus specimens). Sixteen of 78 patients (20.5%) were infected with two or three H. pylori strains with different genotypes. However, no statistically significant differences among cagA occurrence, the different vacA subtypes and PU or CG could be found (each P>0.05). Similarities of the nucleotide sequences from vacA gene s region PCR products of six isolates and from vacA gene m region PCR products of four isolates were 93.2% to 98.3% and 93.8% to 97.6%, respectively, compared to the reported corresponding sequences.Conclusions The dominant genotypes of H. pylori in PU or CG patients in Zhejiang area may be cagA+ s1a/m2 and cagA+ s1a/m1b. Numerous coinfections with different H. pylori strains in PU or CG patients indicate diversity of the infected H. pylori origins. s and m regions of vacA gene from different H. pylori isolates show high nucleotide sequence similarities. cagA gene positive rate, different vacA gene subtypes and coinfection with different H. pylori strains are not closely associated with severity of the diseases.     

Identification and distribution of the clinical isolates of imipenem-resistant Pseudomonas aeruginosa carrying metallo-β-lactamase and/or class 1 integron genes

To investigate the distribution of the genes of two major metallo-β-1actamases （MBL;i.e.,IMP and VIM） and class 1 integrons （intI） in the clinical imipenem-resistant Pseudomonas aeruginosa, a total of 65 isolates, from a university hospital in Sichuan between December 2004 and April 2005 were screened for MBL genes by PCR using primers specific for blaIMP-1, blaVIM and blaVIM-2 genes. The MBL-positive isolates were further assessed for class 1 integrons by PCRusing specific primers. The nucleotide sequences of several PCR products were also determined. The results revealed that the blaVIM gene was found in 81.5% （53/65） of all isolates, blaVIM-2 gene was found in only 1 isolate and the intl gene was observed in 45.3% （24/53） of blaVIM-positive isolates. One isolate carried simultaneously both blaIMP-1 and intl genes, and to the best of our knowledge this is the first report of such isolate in southwest China. These observations highlight that the genes for VIM β-1actamase and class 1 integrons were predominantly present among the imipenem-resistant P. aeruginosa tested, confirming the current widespread threat of imipenem-resistant, integron-borne P.aeruginosa.     

1989～1999年我院烧伤病原菌和细菌耐药性变迁

目的:通过对我院1989～1999年烧伤感染病原菌及耐药性的调查,分析烧伤感染病原菌和细菌耐药性变迁。方法:对1989年6月～1994年5月及1994年6月～1999年6月两个时期的烧伤病人,分别进行血培养和创面培养细菌检出的情况及药敏调。结果:血培养结果表明,铜绿假单胞菌上升至第1位;占24.2％;金黄色葡萄球菌退居第3位,其中,G~4球菌占56.5％,G杆菌占30.1％。创面培养,绿脓杆菌居首,占19.4％;阴沟杆菌居第2位,占14.1％,其中G杆菌占58.4％,G~4球菌占35.3％。阴沟杆菌,嗜麦芽黄单胞菌,粪肠球菌,以及白色念珠菌有明显上升。结论:烧伤感染病原菌和细菌耐药性变迁与头孢他啶,亚胺培南等广谱抗生素广泛应用有一定关系。     

570株铜绿假单胞菌对11种抗菌药物的敏感性及其耐氟喹诺酮机制

目的　了解铜绿假单胞菌的耐药性及其耐氟喹诺酮的机制。方法　Kirby Bauer琼脂扩散法测定 5 70株铜绿假单胞菌对 11种抗菌药物的敏感性 ,琼脂稀释法测定其中 111株对两种氟喹诺酮的最低抑菌浓度 (MIC)。直接序列分析和聚合酶链反应 限制性长度多态性 (PCR RFLP)分析环丙沙星耐药菌株gyrA和parC基因突变。结果　 5 70株铜绿假单胞菌对头孢吡肟、亚胺培南、阿米卡星和头孢他啶的敏感性较高 ,其耐药率分别为 10 9%、11 1%、11 8%、12 5 %。环丙沙星、头孢哌酮 /舒巴坦和哌拉西林的耐药率分别为 2 9 2 %、2 3 5 %和 33 7%。耐环丙沙星和ICU临床分离的菌株对所测试的抗菌药物的耐药率均较高 ,6 0 %以上为多重耐药 (MDR)菌株。 80株耐环丙沙星菌株中 6 6株( 75 % )出现gyrA基因Thr83(ACC)→Ile(ATC)突变 ,5 2株 ( 6 5 % )发生parC基因Ser87(TCG)→Leu(TTG)突变 ,同时存在上述两种突变的有 5 2株 ( 6 5 % ) ,而 31株环丙沙星敏感菌中未发现上述突变。gyrA和parC双基因突变对环丙沙星的MIC显著高于单独gyrA基因突变株和不存在上述突变的菌株 ,P <0 0 5。结论　铜绿假单胞菌的耐药性日趋严重 ,特别是耐环丙沙星和ICU分离株对多种抗菌药物耐药 ;氟喹诺酮类药物作用靶位gyrA和parC的基因突变是临床分离铜绿假单     

实时荧光核酸恒温扩增技术在淋球菌检测中的应用分析

目的采用统计学方法评价实时荧光核酸恒温扩增技术(SAT)在临床淋球菌检测中的应用。方法以淋球菌培养法为标准对照,采用实时荧光核酸恒温扩增技术同时对150例疑似患者的生殖道拭子标本和尿液标本进行检测,对结果进行统计学分析比较,并用PCR法进行验证。结果使用SAT检测的生殖道拭子和尿液阳性检出均为110例,培养法阳性检出为108例,其中培养法检出阴性的2例通过PCR验证为阳性。两种取样方式的SAT检测法和培养法三者阳性率差异均无统计学意义(P0.05);以培养法为金标准,SAT检测拭子的敏感度为100.0%,特异度为95.2%;SAT检测尿液标本的敏感度为100.0%,特异度为95.2%。结论采用实时荧光核酸恒温扩增技术对生殖道拭子和尿道标本的检测效果与培养法相比,阳性率高,敏感度较高,且尿道标本收集方法比较简便,患者依从性好,可代替拭子道取样方法,此方法可在临床推广应用。     

DETECTION OF PATHOGENS CAUSING GENITAL ULCER DISEASE BY MULTIPLEX POLYMERASE CHAIN REACTION

Objective To establish a multiplex polymerase chain reaction （M-PCR） assay for simultaneous detection of pathogens causing genital ulcer disease （GUD）. Mothods Based on the gene-specific region of the following pathogens： Chlamydia trachomatis omp l/ompb, herpes simplex virus （HSV） DNA polymerase, Treponema pollidum tpp47, Haemophilus ducreyi 16s rRNA, four sets of primers were designed and an M-PCR assay was developed to detect four pathogens in one test. The assay was evaluated with diagnostic result of golden standard for each pathogen. Results Of the 51 clinical samples, M-PCR showed slightly higher positive rate （47.1%） of HSV than cell culture （23.6%）. Meanwhile, the positive rate of T. pallidum detected by M-PCR and dark-field microscopy was 19.6% （10/51） and 15.7% （8/51）, respectively. Only one sample was positive for H. ducreyi and no sample was positive for C. trachomatis detected by both M-PCR assay and culture. Conclusion This primary study indicated that M-PCR assay can simultaneously and rapidly detect the four etiologic pathogens causing GUD.     

耐亚胺培南铜绿假单胞菌的β-内酰胺类耐药基因研究

[目的]了解临床分离的耐亚胺培南铜绿假单胞菌（PAE）的相关耐药基因存在状况。[方法]对16株铜绿假单胞菌采用PCR检测亚胺培南耐药的相关基因（TEM、OXA-10群、SHV、oprD2、IMP、CARB、blaDHA、VIM、PER、GES、VEB）。[结果]16株铜绿假单胞菌TEM阳性4株（25.00%）、OXA-10群阳性5株（31.25%）、SHV阳性5株（31.25%）、orpD2阳性16株（100%）、IMP阳性2株（12.50%）CARB阳性1株（6.30%）、blaDHA阳性1株（6.30%）、VIM阳性1株（6.30%）、PER阳性0株（0）、GES阳性1株（6.30%）、VEB阳性0株（0）,且部分菌株携带3个以上的耐药基因。[结论]临床分离的耐亚胺培南铜绿假单胞菌携带多种耐药基因,没有导致院内感染的流行耐药菌株。     

Polymerase chain reaction for the rapid diagnosis of tuberculous meningitis

Polymerase chain reaction (PCR) based on the amplification of a 169 bp DNA fragment specific for the Mycobacterium tuberculosis complex was evaluated for the rapid diagnosis of tuberculous meningitis (TBM). A total of 105 CSF specimens from clinically suspected cases of TBM were studied. Clinical details of the cases and cytochemical parameters of the CSF specimens were recorded. For PCR 10 CSF specimens from cases other than TBM, 4 non-mycobacterial culture isolates (one strain of E coli, one strain of proteus species and 2 strains of salmonella species) and one sample of sterile distilled water were processed as negative controls. For positive control standard culture of Mycobacterium tuberculosis H37Rv was processed with every batch of specimens. Besides PCR, smear for AFB by the Ziehl-Neelsen (ZN) and the fluorochrome method and culture on Lowenstein-Jensen medium was also carried out. By PCR, 31.42% specimens were found positive, whereas by conventional culture on Lowenstein-Jensen medium only 3.8% specimens were positive.     

铜绿假单胞菌亚胺培南异质性耐药的机制

目的: 探讨铜绿假单胞菌（Pseudomonas aeruginosa,P.aeruginosa）亚胺培南异质性耐药的机制。方法: 分别采用常规的KB法和MIC法检测310株铜绿假单胞菌对亚胺培南的敏感性,双纸片协同试验检测金属β 内酰胺酶,实时定量PCR实验检测外排泵基因MexAB和MexCD的表达水平,半定量结晶紫染色法检测生物膜的形成量,PCR扩增法检测细菌膜孔蛋白OPRD2基因是否缺失。结果： 在临床分离的310株铜绿假单胞菌中,检出244株亚胺培南敏感菌株和66株非敏感菌株,244株中有36株铜绿假单胞菌对亚胺培南具有异质性耐药,总体检出率为11.6%。36株异质性耐药铜绿假单胞菌均不产生金属β 内酰胺酶,但高表达MexAB外排泵,且生物膜的形成量也较高,其中6株检出膜孔蛋白OPRD２基因缺失。 结论： 临床存在部分异质性耐药的铜绿假单胞菌,它们通过形成生物膜、高表达多重耐药外排泵基因和基因缺失等机制介导耐药。     

仅黏菌素敏感的多重耐药铜绿假单胞菌分子流行病学研究

目的从基因水平了解多重耐药铜绿假单胞菌中仅黏菌素敏感铜绿假单胞菌(COS-PA)的流行情况。方法①收集本院灼伤科2001年3月至2003年12月临床分离的铜绿假单胞菌52株,其中COS-PA 32株。②收集外科重症监护病房(SICU)2003年1月至2003年12月临床分离的铜绿假单胞菌57株,其中COS-PA 22株;收集SICU水池下水口非仅黏菌素敏感铜绿假单胞菌(non-COS-PA)1株。③采用多种引物随机扩增DNA多态性(RAPD)技术和统计学方法分析其流行情况。结果①灼伤科2001~2003这3年的COS-PA,经RAPD(引物ER IC2、325、272)分型均为同一型别(Burn-A型)。②SICU 2003年收集的COS-PA,经RAPD(引物325)分型为同一型别(SICU-A型);在SICU收集的non-COS-PA也多数呈同一型别(SICU-B型),自SICU水池下水口收集到的1株non-COS-PA也属于此型。③灼伤科及SICU经RAPD(引物325)联合分析证实,两个科室的COS-PA为不同型别,无同源性。结论灼伤科和SICU都存在科室内COS-PA流行,SICU还存在non-COS-PA流行。应严格消毒隔离措施,加强对铜绿假单胞菌在潮湿环境及患者中流行情况的严密监测。多种引物RAPD分析是判断铜绿假单胞菌流行情况的有利工具。     

16SrRNA基因聚合酶链反应快速检测烧伤脓毒症细菌病原体的研究

目的为早期诊断烧伤脓毒症，探索一种能迅速检测细菌感染的方法。方法按标准收集可疑脓毒症患者39人外周静脉血标本共41份。将每份标本一分为二分别行血培养和用细菌通用引物行16SrRNA基因聚合酶链反应检测，用琼脂糖凝胶电泳分析PCR产物，比较两种方法在检测细菌病原体时的快速性和敏感性，分析血培养及药敏结果。结果16SrRNA基因PCR方法的阳性率（41．16％）是血培养阳性率（21．95％）的近两倍。P〈0．05。16SrRANA基因PCR方法出结果需4、5h，血培养需12～24h（多数需2、3d）。9例血培养阳性患者中，8例为单一铜绿假单孢菌感染，1例为单一溶血链球菌感染。结论16SrRNA基因PCR检测细菌感染的方法具有特异性、快速性、敏感性的优点。在该中心，使用抗生素治疗后，引起脓毒症的细菌主要为铜绿假单孢菌。该菌对目前常用广谱抗生素耐药。     

胆汁细菌学检验及其对抗生素的耐药情况

为了解胆道感染性疾病患者细菌感染的耐药情况,对1995年6月～1998年12月间外科住院患者胆汁进行了培养,并对阳性的菌群分布及其抗生素的耐药状况进行了分析(不包括厌氧菌)。结果发现标本培养阳性率为69．76％(143／205),其中大肠埃希氏菌、铜绿假单胞菌占大多数,分别为31．74％和18．56％。其次,肠杆菌属、肺炎克雷伯氏菌和肠球菌属也较为常见。所有病原菌对常用抗生素均产生了不同程度的耐药性。     

环介导等温扩增技术检测铜绿假单胞菌的研究

用环介导等温扩增技术(LAMP)快速、灵敏地检测铜绿假单胞菌.利用铜绿假单胞菌gbca基因,特异性设计3对LAMP引物,在反应体系中添加羟基萘酚蓝(HNB)荧光染料,通过反应前后颜色的变化肉眼观察结果,并用琼脂糖凝胶电泳加以验证.对临床标本利用LAMP和PCR法平行检测,验证该法灵敏度、特异性.针对铜绿假单胞菌DNA特异性引物的LAMP法可以扩增,其他无扩增产物.本实验建立的LAMP法具备简便、快速、特异性强和灵敏度高的特点,适于实际应用.