Efficacy of Pudilan Xiaoyan Oral Liquid in the treatment of pneumonia induced by drug-resistant Pseudomonas aeruginosa

Background:Pudilan Xiaoyan Oral Liquid(PDL)is a Chinese patent medicine with notable pharmacological properties,including anti-inflammatory and antibacterial effects.Drug-resistant Pseudomonas aeruginosa infection is a common and refractory bacterial infection in clinical practice.Due to its high drug resistance,it brings great challenges to treatment.This study aimed to assess the therapeutic efficacy of PDL in a murine model of pneumonia induced by drug-resistant Pseudomonas aeruginosa.Methods:Three different doses of PDL(11 mL/kg/d,5.5 mL/kg/d,2.75 mL/kg/d)were used to observe lung tissue pathology and inflammatory cytokine levels in pneumonia mouse models induced by multidrug-resistant Pseudomonas aeruginosa(MDR-PA).Additionally,the protective efficacy of PDL against mortality in infected mice was evaluated using a death model caused by MDR-PA.Finally sub-MIC concentration of levofloxacin was used to induce drug-resistant mice pneumonia model to evaluate the role of PDL in reversing drug resistance.Experimental data are expressed as mean±standard deviation.Statistical significance was determined by one-way analysis of variance followed by Tukey’s multiple-comparisons test.Results:Treatment effect of PDL on MDR-PA pneumonia:the medium and small doses of PDL can significantly reduce the lung index of multi-drug resistant bacteria infected pneumonia model mice(P<0.05),the lung index inhibition rates for these groups were 55.09%and 58.43%,and improve the degree of lung tissue lesions of mice;The expression of serum cytokines keratinocyte chemoattractant,tumor necrosis factor-αand monocyte chemoattractant protein-1 could be decreased in the three dosage groups of PDL(P<0.01).PDL treatment not only lowered the mortality but also extended the survival duration in mice infected with MDR-PA.It was found after sub-MIC concentration of levofloxacin induced resistance of Pseudomonas aeruginosa to pneumonia in mice.Compared with the model group,the lung index of mice in high and medium PDL doses was significantly reduced(P<0.05),with inhibition rates of 32.16%and 37.73%,respectively.Conclusion:PDL demonstrates protective effects against MDR-PA infection pneumonia,notably decreasing serum inflammatory factor levels.It shows promise in mitigating antibiotic resistance and offers potential for treating pneumonia resulting from Pseudomonas aeruginosa resistance.

Study on the Effect of Shuanghuanglian Powder Needle Combined with Cefoperazone and Sulbactam Sodium on Pseudomonas aeruginosa in Vitro

Objective: To explore the antibacterial activity of combined use of Shuanghuanglian and cefoperazone sulbactam sodium on resistant strains of Pseudomonas aeruginosa. Methods: The Pseudomonas aeruginosa strains which were sensitive and resistant to cefoperazone sulbactam sodium were selected to prepare different test bacterial solutions respectively;The experimental liquid of Shuanghuanglian and Cefoperazone Sulbactam Sodium were prepared separately and set as different test groups and control groups;The Drug Sensitivity Tests of Shuanghuanglian and cefoperazone sulbactam sodium at different concentration gradients which were used alone or used in combination were carried out for different strains with sensitivity and resistance, And use standard entry as a reference control. Result: The results of drug sensitivity test of Shuanghuanglian combined with Cefoperazone-Sulbactam sodium against the resistant strains of Pseudomonas aeruginosa were compared with the results of drug sensitivity test of the two separately used, and the difference was statistically significant (P 〈 0.05) [The drug sensitivity test results of Shuanghuanglian and cefoperazone sulbactam sodium to Pseudomonas aeruginosa resistant strains were statistically significant compared with the drug sensitivity test results of Shuanghuanglian and Cefoperazone Sulbactam Sodium used separately (P 〈 0.05)];There was a dependence between strains and concentration in the effect of the combination of the two drugs. Conclusion: The combination of Shuanghuanglian and cefoperazone sulbactam sodium has synergistic antibacterial or bactericidal effect on Pseudomonas aeruginosa resistant strains. .

Clinical Distribution and Drug Resistance of 224 Strains of Pseudomonas Aeruginosa

Objective:To provide evidence for a rational and effective prevention and treatment of Pseudomonas aeruginosa,the clinical characteristics and the resistance to various antibiotics of were investigated.Methods:A retrospective analysis of 224 strains of Pseudomonas aeruginosa isolated from various specimens from various clinical departments of our hospital(April 1,2018 to June 31,2019)were conducted.Identification and drug susceptibility test of isolated strains was performed using a fully automatic bacterial identification analyzer(MicroScan WalkAway-96 plus),and data analysis was performed using WH0NET5.6 software.Results:Among all the bacteria isolated in our hospital during the above period,Pseudomonas aeruginosa accounted for 10.09% of them all and 12.57% of Gram-negative bacilli,respectively.These isolates were mainly derived from sputum specimens(68.75%),mainly from male patients(70.54%),and mostly 61-70(27.23%)or 51-60(22.77%)years old.Pseudomonas aeruginosa isolates are mainly from Rehabilitation Ward,ICU,and Liver Transplantation Unit,accounted for 29.91%,12.95% and 10.27% of all isolates,respectively.The sensitivity of Pseudomonas aeruginosa to various antibacterial drugs,in the order of high to low were carbapenems,aztreonam,quinolones,cephalosporins,piperacillin/tazobactam,aminoglycoside,with a lowest resistance rate(2.4%)to amikacin and a highest resistance rate to imipenem(33.0%).Conclusion:The isolation rate of Pseudomonas aeruginosa was relatively stable during the study period,and among all the P.aeruginosa detected,most of them were from the respiratory secretions of elderly male patients.The resistance rate of Pseudomonas aeruginosa isolates to various antibiotics is mainly within 30%.Clinical units such as Rehabilitation Ward,ICU,and Liver Transplantation Unit have a high detection rate;therefore,these departments should be monitored in a focused manner.Our research provides a scientific basis for the rational use of antibiotics and a better control of Pseudomonas aeruginosa infection.

The Study of Effect of Commercial Baicalin Combined with Cefoperazone Sulbactam Sodium on Pseudomonas aeruginosa in Vitro

Objective: To investigate the antibacterial effect of baicalin combined with cefoperazone-sulbactam sodium on drug-resistant strains of Pseudomonas aeruginosa. Method: Pseudomonas aeruginosa strains that are sensitive and resistant to cefoperazone-sulbactam sodium were selected to prepare different test bacterial solutions respectively;The test solutions of baicalin and cefoperazone sulbactam sodium were prepared respectively, and different test groups and control groups were set up;The drug sensitivity tests of different concentration gradients of baicalin and cefoperazone sulbactam sodium used alone and in combination were carried out for different sensitive and drug-resistant strains, and the standard strains were used as parallel control. Result: The drug susceptibility test results of the combined use of baicalin and cefoperazone-sulbactam against Pseudomonas aeruginosa drug-resistant strains were compared with the drug susceptibility results of the two used separately, and the difference was statistically significant (P Pseudomonas aeruginosa.

Preliminary Study on the Antibacterial Activity of Six Medicinal Plants against Two Naso-Pharyngeal Pathogens—Streptococccus pyogenes and Pseudomonas aeruginosa

Objective: The objective is to study the antibacterial activity of six medicinal plants against two naso-pharyngeal pathogens and determination of total phenol contents in ethanol extracts of those plants. Methods: Different serial concentrations (0.05 g/mL, 0.1 g/mL, 0.2 g/mL, 0.4 g/mL) of ethanolic and acetone extracts of Piper nigrum L. (Piperaceae), Ocimum sanctum Linn., Plectranthus amboinicus L. (Lamiaceae), Ayapana triplinervis M.Vahl. (Asteraceae), Cinnamomum zeylanicum L. (Lauraceae), Allium schoenoprasum Linn. (Liliaceace) were evaluated for the antibacterial activity using disc diffusion method against gram positive Streptococcus pyogenes and gram negative Pseudomonas aeruginosa. The extracts were prepared from different parts of the plants. The total phenol content was estimated using folin-ciocaltau reagent in catechol equivalents. Results: Majority of the extracts had inhibitory effect against the tested bacteria at different concentrations. In ethanol extracts, Plectranthus amboinicus exhibited the maximum zone of inhibition (14 mm) at 0.05 g/mL concentration against Streptococcus pyogenes, and Ocimum sanctum showed highest zone of bacterial inhibition (19 mm) at 0.05 g concentration against Pseudomonas aeruginosa. In acetone extracts, Piper nigrum had the maximum zone of bacterial inhibition (17 mm) in 0.4 g/mL concentration against Streptococcus pyogenes and Cinnamomum zeylanicum and Allium schoenoprasum exhibited the highest zone of bacterial inhibition (0.4 g/mL) against Pseudomonas aeruginosa. The ethanol extract of Plectranthus amboinicus contained the highest amount of phenol (0.8 mg/mL) and Allium schoenoprasum contained the lowest amount (0.62 mg/mL). In acetone, Cinnamomum zeylanicum contained highest phenol content (0.78 mg/mL). Conclusion: All these investigations pave way to the molecular modeling of the lead phyto compounds present in the studied plants, and also in finding out their biochemical action in various metabolic pathways and reactions of infection.

某院铜绿假单胞菌的临床分布及耐药性分析

目的统计大连市友谊医院近六年来铜绿假单胞菌感染的菌株来源与临床分布,分析11种临床常见抗菌药的耐药情况,为临床治疗铜绿假单胞菌感染及合理使用抗菌药物提供理论依据。方法回顾性分析我院2017年1月-2022年12月铜绿假单胞菌感染的临床送检标本1316株,进行细菌体外分离培养与药敏实验,应用Whonet 5.6软件分析药敏结果,Spss 23软件对6年内该菌耐药率的变化进行卡方检验。结果1316株铜绿假单胞菌中,主要来源于痰液907株(68.9%)和尿液167株(12.7%)。这些菌株主要分布于重症医学科513株(39.0%)和老年病科359株(27.3%)。在所统计的11种抗生素中,哌拉西林/他唑巴坦、头孢吡肟和氨曲南三种药物6年内耐药率变化显著,具有统计学意义(P<0.05);其间耐药率最高的是2021年左氧氟沙星(37.0%),耐药率最低的是2022年头孢吡肟(8.3%)。结论本院铜绿假单胞菌的碳青霉烯类和喹诺酮类耐药率较高,哌拉西林/他唑巴坦呈耐药率逐年上升;微生物室应加强耐药菌监测的同时,临床也要加强抗菌药物的管理。

2020—2022年某医院主要细菌耐药情况分析

目的通过分析福建中医药大学附属第二人民医院2020年1月—2022年12月常见细菌的耐药性,了解医院的细菌耐药情况,为临床提供合理使用抗菌药物的参考依据。方法通过回顾性分析方法对2020年1月—2022年12月福建中医药大学附属第二人民医院患者标本分离的病原菌株进行统计,从病原菌标本分布、病原菌种类及耐药等方面进行分析。结果2020—2022年分离的菌株共计12009株,呼吸道标本(包括痰、肺泡灌洗液)最多。前5位为大肠埃希菌2139株(17.81%)、肺炎克雷伯菌1342株(11.17%)、铜绿假单胞菌1094株(9.11%)、金黄色葡萄球菌为主793株(6.60%)、鲍曼不动杆菌541株(4.50%)。耐碳青霉烯类肺炎克雷伯菌(Carbapenemresistant K.pneumoniae,CR-KP)和耐碳青霉烯类铜绿假单胞菌(Carbapenemresistant Pseudomonas aeruginosa,CR-PA)2022年比2020年明显升高,差异有统计学意义(P<0.01)。其余3种耐甲氧西林金黄色葡萄球菌(Methicillin-resistant S.aureus,MRSA)、耐碳青霉烯类大肠埃希菌(Carbapenemresistant Escherichia coli,CR-EC)、耐碳青霉烯类鲍曼不动杆菌(Carbapenemresistant Acinetobacter baumannii,CRAB)3年间检出率差异无统计学意义(P>0.05)。鲍曼不动杆菌对大多数常用抗菌药物耐药率>50%。结论福建中医药大学附属第二人民医院主要以革兰阴性杆菌为主,鲍曼不动杆菌对大多数常用抗菌药物耐药率>50%,CR-KP(耐碳青霉烯类肺炎克雷伯菌)和CR-PA(耐碳青霉烯类铜绿假单胞菌)的耐药率呈增高趋势,建议临床密切关注病原菌耐药情况的变化,减少多重耐药菌的产生。

2016~2022年昆明市临床分离铜绿假单胞菌耐药性变迁及交叉耐药表型分析

目的了解该院七年来铜绿假单胞菌耐药率变化及耐药表型特征,为铜绿假单胞菌感染早期经验治疗提供依据,并为后期耐药机制研究奠定一定的基础。方法使用Whonet 5.6软件按年统计昆明市第一人民医院2016~2022年从临床标本分离并报告的铜绿假单胞菌药物敏感性数据,以亚胺培南、美罗培南、头孢他啶及头孢吡肟等药物为主要观察对象,统计并分析上述药物不同耐药性组合下其它抗生素耐药性情况。结果七年共检出铜绿假单胞菌1920株,其中多黏菌素耐药率最低为1%;阿米卡星、庆大霉素、妥布霉素平均耐药率分别为6.4%,8.6%,5.1%;喹诺酮类与哌拉西林/他唑巴坦、头孢他啶、头孢吡肟耐药率相似,在10%~20%之间;亚胺培南和美罗培南未显示出更优秀的敏感性,耐药率分别为21.3%和18.4%。各主要抗生素耐药率在七年间存在波动,多数自2017年上升,至2018年达峰值后回落,2020年后趋于平稳并与国内平均水平一致;亚胺培南敏感株1393株,该部分美罗培南敏感率为96.2%;美罗培南敏感株1451株,该部分亚胺培南敏感率为89%。亚胺培南、美罗培南均耐药共369株,该部分头孢他啶敏感率为33.1%。β-内酰胺类抗生素耐药种类越多,阿米卡星、环丙沙星敏感率越低。结论该院铜绿假单胞菌药物敏感性总体较好,应坚持严格的抗生素使用管理。氨基糖苷类可作为经验治疗首选药物,喹诺酮类次之,碳青霉烯类和头孢他啶亦为备选。该院铜绿假单胞菌对β-内酰胺类抗生素耐药机制呈多样性,碳青霉烯耐药株可由不同机制分别或共同介导。

广西地区耐碳青霉烯类铜绿假单胞菌的流行特征、耐消毒剂基因检测及同源性分析

目的了解耐碳青霉烯类铜绿假单胞菌(CRPA)临床分离菌株的流行特征,检测、分析耐消毒剂基因携带情况及菌株同源性。方法回顾性分析该院2020—2022年临床分离得到的252株CRPA的临床分布特点、耐药趋势。并随机选取其中30株菌株,采用聚合酶链反应检测耐消毒剂基因qacEΔ1-sul1及aceⅠ的表达情况;采用肠杆菌科基因间重复序列-聚合酶链反应检测菌株同源性。结果CRPA的标本来源主要为痰液(63.49%)、肺泡灌洗液(17.06%);来自综合重症监护病房(ICU)和其他ICU占比为40.87%,来自非ICU的占比为59.13%,非ICU病区主要分布在康复医学科(14.68%)和呼吸科(12.70%)。除头孢吡肟、头孢他啶/阿维巴坦、阿米卡星、庆大霉素、妥布霉素、多黏菌素外,CRPA对其他临床常用抗菌药物的耐药率均在30.00%以上。CRPA在ICU及非ICU病区的耐药率比较,差异无统计学意义(P>0.05)。30株临床分离的CRPA qacEΔ1-sul1和aceⅠ基因检出率分别为16.67%、26.67%。同源性分析结果显示A型和B型为主要流行株。结论CRPA对常用抗菌药物普遍耐药,应加强对CRPA菌株的耐药性监测,根据药敏试验结果合理选择抗菌药物,减少耐药株的产生。CRPA中qacEΔ1-sul1和aceⅠ两种耐消毒剂基因均有检出,但检出率均较低,实际工作中应加强对各病区的消毒效果监测。CRPA中存在A、B两种优势型别的克隆流行,应作为铜绿假单胞菌院内感染防控的重点。

医院2019—2023年铜绿假单胞菌临床分布及耐药性分析

目的分析医院2019—2023年铜绿假单胞菌临床分布及耐药性。方法选取2019—2023年西安市第三医院收治的住院患者及门诊送检的各类标本,获取出490株铜绿假单胞菌,对该菌进行药敏试验,分析标本来源及科室分布,并统计铜绿假单胞菌对常见抗菌药物的耐药性。结果490株铜绿假单胞菌中,痰液检出率最高,占63.27%,其次为尿液,占13.67%;呼吸科检出率最高,占24.69%,其次为ICU、神经外科,分别占18.78%、11.02%。铜绿假单胞菌对头孢曲松完全耐药,对阿米卡星、环丙沙星、美罗培南、庆大霉素、头孢吡肟、妥布霉素、亚胺培南、左氧氟沙星、头孢哌酮/舒巴坦、哌拉西林/他唑巴坦、头孢他啶的敏感率均>80.0%。结论医院2019—2023年铜绿假单胞菌多来源于痰液,主要存在于呼吸科,且铜绿假单胞菌对抗菌药物的整体耐药率偏低。

扶正透邪解毒化瘀方对不同时间点老年肺炎大鼠炎症及免疫功能的影响

目的:探讨扶正透邪解毒化瘀方对多重耐药铜绿假单胞菌肺炎老年大鼠炎症反应与免疫功能的态影响。方法:将实验老年大鼠随机分为空白对照组、模型对照组、西药对照组、中药组、中药提前给药组、中西医结合观察组,动态观察不同时间点各组老年大鼠炎症指标的变化及淋巴细胞增殖水平的变化。结果:中药提前给药组及中西医结合观察组疗效显著,中药组对感染早期的炎症水平改善明显;中西医结合观察组可使T淋巴细胞增殖能力趋于正常;中药提前给药组、中西医结合观察组能抑制感染后B淋巴细胞的增殖并使其趋于正常。结论:扶正透邪解毒化瘀方能够调节感染早期老年大鼠机体过度的免疫炎症反应,中西医结合治疗能够纠正多重耐药铜绿假单胞菌感染后紊乱的免疫状态,中药提前给药能够对多重耐药铜绿假单胞菌感染后的炎症反应及免疫紊乱起到一定的预防及治疗作用。

铜绿假单胞菌耐药机制及治疗技术研究新进展

铜绿假单胞菌(PA)是最常见的条件致病菌之一,严重危害人类的健康。随着广谱抗生素的大量应用,导致多重耐药菌株的出现,从而使治疗更加棘手。研究发现,免疫力低下的人群容易诱发PA的感染,随着临床耐药PA检出率的增加及抗生素的治疗效果减弱,部分患者的生活质量及预后受到很大影响。PA的感染难以控制的原因是复杂的耐药机制,其中包括生物膜的形成、孔蛋白的介导作用、外排泵以及抗菌灭活酶的产生等。由于抗生素的应用不能有效控制PA的感染,所以针对耐药机制在药物治疗方面进行研究,其中包括外排泵抑制剂、抗生素佐剂、相关灭活酶抑制剂、外膜透化剂。此外,噬菌体疗法、中药疗法、一氧化氮疗法及纳米技术等特殊治疗也展现出良好的治疗效果,疫苗领域及增强自身免疫力也是未来预防PA感染的重要研究方向。因此,研究PA的耐药机制及治疗技术对PA感染的预防和治疗具有深远的意义。

铜绿假单胞菌前噬菌体的生物信息学分析

目的预测分析83株铜绿假单胞菌中前噬菌体携带情况和分布特征,以及鉴定前噬菌体编码的抗生素耐药基因和毒力因子。方法采用Phaster在线工具对菌株基因组序列进行前噬菌体预测,采用Cadr数据库和Vfdb数据库进行耐药基因和毒力基因注释分析。结果83株铜绿假单胞菌中共检测出601条前噬菌体序列,其中完整型304条,不完整型161条,可疑型136条,平均每株铜绿假单胞菌中至少存在3条完整的前噬菌体。平均每株铜绿假单胞菌中携带的所有前噬菌体占其基因组长度比例约为2%~4%。61条前噬菌体预测到125个耐药基因,分别来自23个不同家族,产生7种耐药机制。356条前噬菌体共编码545个毒力基因,归类为119种毒力基因,编码78种相关毒力因子,平均每个前噬菌体携带1~2个毒力因子编码基因,少数前噬菌体携带3条及以上。毒力因子的宿主来源除了铜绿假单胞菌外,百日咳杆菌、微黄分枝杆菌、鸟分枝杆菌及其亚种、肺炎克雷伯菌及其亚种也占有较高来源比例。结论前噬菌体在铜绿假单胞菌中分布广泛且携带多种耐药和毒力基因,对铜绿假单胞菌耐药性获得、传播和致病性演变具有重要影响。

耐碳青霉烯类铜绿假单胞菌毒力基因的表达与耐药性研究

目的探讨临床检测的耐碳青霉烯类铜绿假单胞菌中Ⅲ型分泌系统(T3SS)4种毒力基因(exoT、exoY、exoS、exoU)的表达情况及其对抗菌药物的耐药性。方法收集安徽医科大学附属安庆第一人民医院2022-2023年临床检测的各种来源的耐碳青霉烯类铜绿假单胞菌87株,采用Vitek2-Compact仪器对菌株进行鉴定,并且进行药敏试验,采用聚合酶链反应对T3SS的4种毒力基因(exoT、exoY、exoS、exoU)进行检测,比较不同毒力基因型组合与抗菌药物耐药性之间的关系。结果87株耐碳青霉烯类铜绿假单胞菌表达exoT与exoY基因的阳性率均为100.00%(87/87),exoS基因阳性率为77.01%(67/87),exoU基因阳性率为27.59%(24/87)。共检出4种类型的T3SS毒力基因型组合,以exoT+/exoY+/exoS+/exoU-基因型组合为主,阳性率为71.26%(62/87),其次是exoT+/exoY+/exoS-/exoU+基因型组合,阳性率为21.84%(19/87)。药敏试验结果显示,exoT+/exoY+/exoS-/exoU+基因型组合对大部分常见抗菌药物,如阿米卡星、妥布霉素、头孢吡肟、头孢他啶、头孢哌酮/舒巴坦、环丙沙星、哌拉西林的耐药率均明显高于exoT+/exoY+/exoS+/exoU-基因型组合,差异均有统计学意义(P<0.05)。结论该院检出的耐碳青霉烯类铜绿假单胞菌携带表达的T3SS毒力基因主要以exoT+/exoY+/exoS+/exoU-基因型组合为主,不同毒力基因型组合的耐碳青霉烯类铜绿假单胞菌对常见抗菌药物的耐药性存在差异。

隐丹参酮对铜绿假单胞菌的生长及生物膜形成的抑制作用

为研究隐丹参酮(cryptotanshinone)对铜绿假单胞菌(Pseudomonas aeruginosa)的生长和生物膜形成的影响及其机制,采用微量稀释法测定隐丹参酮对铜绿假单胞菌的最小抑菌质量浓度(MIC),利用结晶紫染色法、倒置显微镜和激光扫描共聚焦显微镜观察隐丹参酮对铜绿假单胞菌生物膜形成的影响,再采用实时荧光定量聚合酶链反应(RT-qPCR)检测隐丹参酮对铜绿假单胞菌生物膜形成相关基因LasI、LasR、RhlI和RhlR的表达的影响。隐丹参酮对铜绿假单胞菌的MIC为800.0μg/mL;结晶紫染色后,隐丹参酮的质量浓度为800.0μg/mL时抑制菌膜效果最佳,且呈剂量依赖性(P<0.01);利用激光共聚焦显微镜观察隐丹参酮对生物膜的形成和分布,200.0μg/mL时抑菌效果显著提升。在200.0~800.0μg/mL的质量浓度范围内,800.0μg/mL的隐丹参酮对LasI、LasR、RhlI和RhlR基因的表达具有抑制作用(P<0.01)。隐丹参酮抑制铜绿假单胞菌生物膜形成机制可能是通过下调LasI等基因的表达。本研究表明,隐丹参酮可以作为群体感应抑制剂干预铜绿假单胞菌的耐药,为临床多重耐药铜绿假单胞菌(MDR-PA)的治疗提供参考。

耐碳青霉烯类铜绿假单胞菌的耐药机制及特征分析

目的探究本院耐碳青霉烯类铜绿假单胞菌(carbopenem-resistant pseudomonas aeruginosa,CRPA)的耐药机制及特征。方法用BD phoenix M50全自动鉴定药敏分析仪对菌株进行鉴定及药敏分析,微量肉汤稀释法测定头孢他啶/阿维巴坦的最低抑菌浓度(minimal inhibitory concentration,MIC),采用改良碳青霉烯类失活法(modified carbapenem inactivation method,mCIM)和胶体金免疫层析法对菌株进行碳青霉烯酶表型检测,采用全基因组测序对筛查阳性菌株进行碳青霉烯酶耐药基因检测及ST分型。结果①收集22株临床分离CRPA,其中耐药率最低的抗菌药物为头孢他啶/阿维巴坦(22.7%),然后依次为庆大霉素和阿米卡星(27.3%),哌拉西林/他唑巴坦(59.09%),头孢吡肟(63.6%),头孢他啶和氨曲南(77.27%),环丙沙星(86.36%),左氧氟沙星(95.45%)。②22株CRPA表型检测结果显示产碳青霉烯酶共5株(22.7%)。③全基因组测序结果显示,有4株ST549型CRPA携带金属β-内酰胺酶IMP-45及丝氨酸β-内酰胺酶OXA-1,OXA-50,1株为ST245型CRPA携带金属β-内酰胺酶NDM-1,即5株CRPA菌株均产金属β-内酰胺酶。结论产碳青霉烯酶不是本院CRPA对碳青霉烯耐药主要机制,CRPA对头孢他啶/阿维巴坦耐药率低且其耐药机制主要是产金属β-内酰胺酶。

老年医院分离铜绿假单胞菌的耐药性及耐消毒剂基因分析

目的分析老年患者分离铜绿假单胞菌(Pseudomonas aeruginosa,PA)的耐药性和耐消毒剂基因的分布情况。方法收集2019—2022年老年患者分离的111株PA,采用VITEK 2 Compact进行细菌鉴定和耐药性分析,PCR法检测耐消毒剂基因,采用SPSS 21.0对结果进行统计分析。结果15种药物中PA耐药率最高的是左氧氟沙星(30.63%),最低的是多黏菌素B(1.80%),耐碳青霉烯铜绿假单胞菌(carbapenem-resistant Pseudomonas aeruginosa,CRPA)占比为23.42%(26/111),多重耐药铜绿假单胞菌(multidrug-resistant PA,MDR-PA)占比为20.72%(23/111)。共检出3种耐消毒剂基因:qacE⊿1-sulΙ、emrE及merA基因,阳性率分别为16.22%(18/111)、91.89%(102/111)和46.85%(52/111),只有3株PA所有消毒剂基因均未检出。消毒剂基因携带模式最多见为模式c(41.44%,46/111);其次为模式f(36.94%,41/111)。CRPA组和非CRPA组之间、MDR组和非MDR组之间qacE⊿1-sulΙ检测阳性率具有显著差异。此外,MDR组qacE⊿1-sulΙ、emrE及merA基因同时检出的比率显著高于非MDR组。结论老年患者分离PA中携带耐消毒剂基因的比率高,携带qacE⊿1-sulΙ基因与耐药性之间可能存在一定的相关性。临床需科学合理使用消毒剂和抗菌药物,尽量减少细菌产生对两者的抗性。

树鼩源碳青霉烯耐药性铜绿假单胞菌的分离鉴定及生物学特性研究

【目的】确定养殖过程中死亡树鼩的致病原因并分析病原的生物学特性、致病性和耐药情况,制定防治方法。【方法】对死亡树鼩进行病理剖检,记录肺脏、肝脏、脾脏、肠道等主要脏器的病变情况,通过平板划线法从主要病变脏器中分离单个菌落,观察菌落形态并进行革兰染色镜检,通过生化试验、药敏试验、致病性试验及16S rRNA基因测序和全基因组测序等手段对分离株进行进一步分析。【结果】从死亡滇西树鼩的脾脏、肠道中分离获得1株碳青霉烯耐药性铜绿假单胞菌,分离菌株在LB琼脂培养基中呈现产绿色水溶色素的灰白色菌落,镜检为革兰阴性杆菌;生化试验进一步证实分离菌为非发酵菌;通过序列进化分析可知,分离菌与GenBank收录的土壤来源铜绿假单胞菌F4相似性高达99%。药敏试验结果显示,分离菌对亚胺培南和美罗培南等碳青霉烯类及左氧氟沙星等喹诺酮类抗菌药均具有一定的耐药性。致病性试验结果显示,该分离株对成年昆明小鼠的半数致死量(LD 50)为3.382×108 CFU,对死亡小鼠脏器组织进行病理学观察,可见肺脏、肝脏、脾脏、肠道等的炎性损伤和坏死。对各脏器再次进行细菌分离,基于16S rRNA序列分析发现铜绿假单胞菌可在死亡小鼠的脾脏、肠道、血液中存在,具有较高的致病性。全基因组测序结果证实,分离菌具有碳青霉烯耐药性相关基因,并与铜绿假单胞菌高致病参考菌株具有相似的毒力因子基因。【结论】本研究从病死滇西树鼩体内分离出碳青霉烯耐药性铜绿假单胞菌,为树鼩微生物致病研究和驯化养殖提供了新的理论依据。

难治性耐药铜绿假单胞菌血流感染的危险因素分析

目的探讨难治性耐药铜绿假单胞菌(DTR-PA)血流感染患者的临床特点和危险因素。方法回顾性分析2017年1月至2022年12月中山大学附属第一医院发生铜绿假单胞菌血流感染的患者病历资料,采用多因素Logistic回归分析探究DTR-PA血流感染的独立危险因素,采用Cox比例风险回归模型探讨铜绿假单胞菌血流感染死亡相关的危险因素。结果共纳入114例铜绿假单胞菌血流感染患者,平均年龄(51.05±17.55)岁,28 d死亡率为14.0%(16/114),DTR-PA共有24株(21.1%,24/114)。年龄、入住ICU、糖尿病、低白蛋白血症、感染性休克与DTR-PA血流感染有关(P<0.05)。多因素Logistic回归分析显示,感染性休克是患者发生DTR-PA血流感染的独立危险因素(P<0.05)。入住ICU是铜绿假单胞菌及DTR-PA血流感染患者死亡的独立危险因素(P<0.05)。结论DTR-PA血流感染的危险因素有多种,感染性休克和入住ICU分别是DTR-PA血流感染和死亡的独立危险因素,临床应重视对该类菌的监测。

多重耐药假单胞菌属细菌对碳青霉烯类和氨基糖苷类抗生素耐药的分子机制研究

【目的】探讨多重耐药假单胞菌属细菌对碳青霉烯类和氨基糖苷类抗生素耐药的分子机制。【方法】选2022年1月至12月于本院检验科收到的来自该医院不同病区的6株多重耐药恶臭假单胞菌和铜绿假单胞菌。采用Vitek-2compact全自动鉴定仪和16s rDNA序列引物鉴定并核实细菌种属。E-test药敏试条法测定菌株对临床抗生素抑菌浓度。PCR扩增和序列对比明确氨基糖苷类和碳青霉烯酶类甲基化酶的基因亚型。【结果】6株假单胞菌属细菌中有4株铜绿假单胞菌、2株恶臭假单胞菌,均为Carba NP阳性,且产碳青霉烯酶;菌株对青霉素类、碳青霉烯类、氨基糖苷类、头孢菌素类、四环素类抗生素均显示为耐药性。6株菌株中,仅SY456为氨曲类耐药,最小抑菌浓度值(MIC值)为92μg/mL,而其他菌株均为氨曲类耐药中介或敏感。Carba NP法筛查出6株菌均产耐药B类碳青酶烯酶。经与overlap PCR序列比对显示,6株菌株携带armA基因和blaIMP-45基因,3株恶臭假单胞菌SY47、SY153和SY434之间的脉冲场凝胶电泳(PFGE)带型差异显著。【结论】本院存在同时携带armA和blaIMP-45的多重耐药恶臭假单胞菌和铜绿假单胞菌,临床医师需根据耐药菌株类型调整用药策略。