Carba NP试验在产碳青霉烯酶肠杆菌科细菌检测价值的meta分析

目的:系统评价Carba NP试验对产碳青霉烯酶菌株的诊断价值.方法:检索PubMed数据库、EMBASE数据库、Cochrane图书馆、维普中文科技期刊数据库、中国知网、万方数据库、中国生物医学文献数据库(CBM),并辅以文献追溯、手工检索,检索时间为2001年1月1日至2016年6月30日.严格按照纳入和排除标准进行文献筛选.参照诊断准确性研究质量评价工具(Quality Assessment of Diagnostic Accuracy Studies 2,UADAS-2)对纳入文献进行质量评价.应用STATA14.0软件进行数据提取及质量评价、统计和数据分析,绘制森林图分析结果,用漏斗图和Deek检验评价纳入文献的发表偏倚情况.结果:纳入的29项研究的合并敏感性和特异性分别是0.97(95％CI:0.93～0.98)和1.0(95％CI:1.0～1.0),说明敏感性及特异性较高.经异质性分析检验,敏感性和特异性的I2分别是96.49(95％CI:95.79～97.18)和69.88(95％CI:58.69～81.08),说明纳入数据存在异质性.结论:Carba NP试验对于产碳青霉烯酶肠杆菌科细菌(Carbapenem-resistant Enterobacteriaceae,CRE)的检测具有很高的敏感性和特异性.     

我院耐碳青霉烯类肠杆菌科细菌碳青霉烯酶不同检测方法的比较

目的 探讨评价碳青霉烯酶的不同检测方法，为实验室检测及临床诊疗提供依据。方法 选用EDTA碳青霉烯灭活方法(mCIM/eCIM)检测耐碳青霉烯类肠杆菌(carbapenem-resistant Enterobacteriaceae, CRE)菌株耐药表型，以两种免疫层析碳青霉烯酶检测试剂盒、荧光定量PCR法检测CRE菌株基因型。统计学方法采用Kappa一致性检验，比较各种检测方法的临床可操作性。结果 经荧光定量PCR扩增后基因测序显示73株肺炎克雷伯菌中71株携带KPC基因，2株携带NDM基因；7株大肠埃希菌中6株携带NDM基因，1株携带KPC基因；mCIM联合eCIM检测KPC的灵敏度为68.1%(49/72),检测NDM的灵敏度为100.0%(8/8);NG-test Carba 5和免疫显色试剂盒检测KPC和NDM灵敏度为100.0%、特异性为100.0%。结论 NG-test Carba 5和免疫显色试剂盒检测可以成为检测我国最常见碳青霉烯酶的快速、方便的诊断工具，从而有助于控制产碳青霉烯酶的分离株在医院间的传播，为院感防控工作起到至关重要的意义。     

儿科患者中对碳青霉烯类不敏感肠杆菌科细菌耐药性和耐药基因的研究

目的 研究儿科临床分离的对碳青霉烯类抗生素不敏感的肠杆菌科细菌耐药性和产生碳青霉烯酶的耐药基因特征.方法 收集2008年1月至2010年12月北京儿童医院住院患儿分离出的46株对碳青霉烯类抗生素不敏感的肠杆菌科细菌.使用琼脂稀释法进行药敏试验,测定抗菌药物的最低抑菌浓度(MIC)值,按照临床实验室标准化研究所(CLSI) 2011年推荐标准判断结果.使用改良Hodge试验和双纸片协同试验,进行产碳青霉烯酶的表型确证.使用PCR方法进行碳青霉烯酶相关耐药基因的检测.采用WHONET5.6软件进行数据分析.结果 46株对碳青霉烯类抗生素不敏感的肠杆菌科细菌,26株为肺炎克雷伯菌,占56.5％,13株为阴沟肠杆菌,占28.3％,7株为大肠埃希菌,占15.2％.对亚胺培南和美罗培南不敏感率分别为肺炎克雷伯菌69.2％和80.8％,阴沟肠杆菌76.9％和100％,大肠埃希菌85.7％和100％.46株肠杆菌科细菌,改良Hodge试验阳性40株(87.0％),双纸片协同试验阳性41株(89.1％).IMP基因阳性38株(82.6％),其余8株均未扩增出特异性条带检测均为阴性.结论 目前儿科临床分离对碳青霉烯类抗生素不敏感菌株中,肺炎克雷伯菌最多,占56.5％.肺炎克雷伯菌、阴沟肠杆菌和大肠埃希菌对碳青霉烯类抗生素亚胺培南不敏感程度低于美罗培南,对碳青霉烯不敏感肠杆菌科细菌主要产生B类金属酶,均为IMP基因型.     

对碳青霉烯类抗生素敏感性下降的肠杆菌科细菌耐药机制的研究及流行病学调查

目的 对临床分离的碳青霉烯类抗生素耐药的肠杆菌科细菌进行耐药机制研究及流行病学调查.方法 收集2010年1月至2010年8月,对碳青霉烯类抗生素敏感性下降的肠杆菌科细菌18株,全自动微生物鉴定仪检测细菌对常见抗生素最低抑菌浓度(MIC),纸片法检测细菌产超广谱β-内酰胺酶、头孢菌素酶、碳青霉烯酶的情况,并用PCR扩增、DNA测序确定所产碳青霉烯酶基因型.脉冲场凝胶电泳对耐药菌进行同源性分析.结果 8株耐药菌全部检出ESBLs酶、AmpC酶,17株检出KPC-2酶,3株EDTA纸片法阳性提示产其他金属碳青霉烯酶,其中两株合并产KPC-2.脉冲场凝胶电泳结果显示15株肺炎克雷伯菌分为6种带型.结论 KPC-2碳青霉烯酶是造成肠杆菌科细菌对碳青霉烯类抗生素敏感性下降的主要原因,并在我院局部短暂流行,携带KPC-2基因的临床菌株同时携带多种耐药基因.     

改良Hodge试验对鲍曼不动杆菌产碳青霉烯酶检测效果的比较

目的比较改良Hodge试验采用三种不同纸片法对耐亚胺培南鲍曼不动杆菌产碳青霉烯酶的检测能力。方法改良Hodge试验采用三种不同纸片检测碳青霉烯酶;采用MicroScan WalkAway 96检测鲍曼不动杆菌的耐药性。结果改良Hodge试验(三种不同纸片法)同时检测33株耐亚胺培南鲍曼不动杆菌,其中应用改良Hodge试验(亚胺培南纸片法),阳性为22株,阳性率为66.7%;改良Hodge试验(厄他培南法),阳性为17株,阳性率为51.5%;改良Hodge试验(美罗培南法),阳性为8株,阳性率为24.2%。结论改良Hodge试验(三种不同纸片法)筛选耐亚胺培南鲍曼不动杆菌产碳青霉烯酶中,改良Hodge试验(亚胺培南纸片法)敏感性最高,而改良Hodge试验(美罗培南纸片法)敏感性最低。     

碳青霉烯类耐药肠杆菌科细菌实验室检测的研究

碳青霉烯类耐药肠杆菌科细菌（CRE）已经成为全球性公共卫生问题,这类细菌往往伴随高致病率、高致残率、高死亡率,为临床治疗带来了极大的挑战。CRE的耐药机制主要是产生碳青霉烯酶。快速、准确地检测产碳青霉烯酶的肠杆菌科细菌,对于合理使用抗生素,预防和控制产酶菌株的传播具有重要意义。本文就实验室检测肠杆菌科细菌产碳青霉烯酶的研究方法进展作一综述。     

一株产碳青霉烯酶IMP-26阴沟肠杆菌的分离鉴定

在过去的几年中,产碳青霉烯酶的肠杆菌科细菌不断被发现,这引起了临床工作者的强烈担忧.我院发现一株碳青霉烯类抗菌药物耐药的阴沟肠杆菌,此临床菌株分离自一位9个月的哮喘患儿,对亚胺培南和厄他培南表现为耐药,其最低抑菌浓度分别为4 μg/ml和16μg／ml.对青霉素类,头孢类及头孢与酶的复合制剂都表现为耐药.进一步的实验显示此菌株的Hodge试验阳性,确定该菌株产碳青霉烯酶.通过对亚胺培南与亚胺培南+EDTA的纸片的抑菌圈直径的比较,发现此种碳青霉烯酶能被EDTA抑制.通过PCR扩增,发现此阴沟肠杆菌含有blaInt11、blaIMP和blaTEM基因,通过测序和比对确定blaIMr的基因型为blaIMP-26,balTEM基因的基因型为blaTEM-104.通过等电聚焦电泳,也证实此阴沟肠杆菌产IMP和TEM酶.     

质粒介导KPC-2型碳青霉烯酶肺炎克雷伯菌儿童分离株耐药基因研究

目的 研究碳青霉烯类耐药肺炎克雷伯菌临床儿童分离株的耐药特点及分子流行病学特征.方法 收集温州医学院附属第二医院2010年7月-2011年6月从儿童标本中分离的耐碳青霉烯类肺炎克雷伯菌12株,所有菌株为非重复菌株,菌种鉴定采用全自动微生物分析仪.改良的Hodge试验筛选产碳青霉烯酶阳性菌株,采用PCR法检测KPC、IMP、bla(s)、VIM、SPM和整合酶基因,测序确定基因型.对菌株进行质粒结合试验、质粒消除试验检测质粒的转移性.脉冲场凝胶电泳(PFGE)分析耐药菌株的同源性.结果 12株耐碳青霉烯类肺炎克雷伯菌对庆大霉素、妥布霉素、阿米卡星、环丙沙星、左氧氟沙星、复方磺胺甲噁唑的敏感率分别为8.3％、41.7％、58.3％、8.3％、8.3％、33.3％;12株菌均携带有KPC-2基因,且同时携带有TEM-1和SHV型β-内酰胺酶基因,其中SHV-11-like和SHV-1 2-like各6株;11株携带CTX-M型基因,其中4株为CTX-M-14-like基因,6株CTX-M-15-like基因;2株携带有OXA-10型基因,1株携带有PER-1基因.未检出NDM-1、GIM、SPM、SIM、VIM型碳青霉烯酶基因.12株均为Ⅰ类整合酶基因(int1)阳性.2株通过接合试验把质粒传递给受体菌EC600.所有接合子blaTEM-1基因阳性、超广谱β-内酰胺酶(ESBL)基因阳性及对亚胺培南、庆大霉素、阿米卡星、妥布霉素和头孢噻肟耐药,接合子ESBL基因型与供菌一致.2株菌经质粒消除后对亚胺培南的MIC值均有较大程度降低,消除后KPC-2基因扩增为阴性.12株KPC-2基因阳性菌株经PFGE分成5个基因型,主要为B型和C型.结论 KPC-2型碳青霉烯酶基因已经在儿童肺炎克雷伯菌中播散,常伴随携带多种类型的ESBL基因和Ⅰ类整合酶基因,部分耐药基因可通过质粒播散.     

重症医学科染KPC-2肺炎克雷伯菌的耐药情况分析及同源性研究

目的重症医学科染KPC-2肺炎克雷伯菌的耐药及同源性情况。方法从2018年4月至2019年2月于我院重症医学科室治疗的患者中提取40例耐碳青霉烯类肺炎克雷伯菌(CRKPN)菌株,对待测菌株进行药敏性试验、mCIM(改良碳青霉烯灭活试验)和eCIM(EDTA改良碳青霉烯灭活试验)联合试验,采用PCR扩增法检测待测菌的耐药基因并进行基因序列检测,采用PFGE对待测菌进行同源性分析。结果在16种抗菌药中,待测菌仅对阿米卡星(27.50%)、庆大霉素(17.50%)、妥布霉素(25.00%)、替加环素(100.00%)以及复方新诺明(55.00%)有敏感性;mCIM和eCIM试验结果显示40株CRKPN均产丝氨酸碳青霉烯酶;PCR试验结果显示待测菌株携带KPC基因阳性率为100%;选择ICU分离的17株CRKPN做PFGE,同源性结果分析显示该病区存在耐碳青霉烯肺炎克雷伯的克隆传播。结论我院重症医学科分离的CRKPN细菌的耐药机制是产丝氨酸碳青霉烯酶,酶基因为KPC-2型,PFGE结果分析显示我院重症医学科存在耐碳青霉烯肺炎克雷伯的克隆传播,需要加强院感监控工作。     

产气肠杆菌中发现碳青霉烯酶KPC-2型

肺炎克雷伯菌碳青霉烯酶(Klebsiella pneumoniae Carbapenemase,KPC),属A类β-内酰胺酶.2010年3月25日我们从一家三等甲等医院老年患者的痰液中分离到1株碳青霉烯类耐药的产气肠杆菌(标本编号:20100323BAA049,经gyrA和parC基因扩增与测序,GenBank比对确认).为了解该菌株的β-内酰胺类耐药机制,我们采用E-test法检测了该菌对26种抗菌药物敏感性,并进行了A类、B类、C类、D类等4类41种β-内酰胺酶基因检测,用改良的三维试验法检测AmpC酶、ESBLs和金属β-内酰胺酶活性,用改良Hodge试验法检测碳青霉烯酶活性.     

Performance of the Rapidec®Carba NP assay for the detection of different carbapenemases in Klebsiella pneumoniae and Escherichia coli strains

PurposeThe spread of infections caused by Enterobacterales strains resistant to carbapenems is a global public health problem, and early detection of carbapenemases is very important to prevent their spread. The rapid detection of carbapenemase production with the new commercial assay Rapidec® Carba NP test is based on the biochemical detection of imipenem hydrolysis. Our study aims to evaluate the performance of the Rapidec® Carba NP test in OXA-48 positive isolates highly prevalent in our country and also in isolates with more than one carbapenemase gene that have an increased prevalence and to examine whether it can be used for confirmation of carbapenemase positivity in the routine laboratory.MethodsA total of 97 strains of 94 carbapenem-resistant Klebsiella pneumoniae and three carbapenem-resistant Escherichia coli isolated from various clinical specimens were included in the study. The results of the Rapidec® Carba NP assay were compared with those obtained by the multiplex PCR test.ResultsThe sensitivity of the Rapidec® Carba NP test was 97.8% for all carbapenemase-positive isolates. Of 90 PCR positive isolates, one OXA-48 and one OXA-48 ?+ ?NDM positive isolates were negative with Rapidec® Carba NP test.ConclusionsThe positive results detected by the Rapidec® Carba NP test make an important contribution to the early detection of carbapenemase production and infection control practices. Since two carbapenemase positive isolates were found to be negative with the Rapidec® Carba NP test in our study, it was concluded that negative results of carbapenem-resistant isolates obtained with this assay should be confirmed with an additional carbapenemase detection method to exclude false-negative results.     

Evaluation of the Rapidec Carba NP Test for Detection of Carbapenemases in Enterobacteriaceae

This study evaluated the performance of the Rapidec Carba NP test, which was introduced recently into the market for the detection of carbapenemase production in a broad spectrum of β-lactamase-producing Enterobacteriaceae clinical isolates. In total, 252 clinical Enterobacteriaceae isolates that had been genetically characterized with respect to carbapenemase, extended-spectrum β-lactamase (ESBL), and AmpC genes were analyzed; 51/252 isolates (20.2%) were genetically confirmed to be carbapenemase producers, whereas 201/252 isolates (79.8%) were genetically negative for the presence of carbapenemase genes. The Rapidec Carba NP test was applied according to the manufacturer''s instructions, and results were read after 30 and 120 min of incubation. The overall sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of the Rapidec Carba NP test were 90.2%, 100%, 100%, and 97.6%, respectively, when the manufacturer''s instructions were followed. Four of 5 false-negative results occurred with OXA-48-like enzymes. After an incubation time of 30 min, the sensitivity was 49%. The sensitivity increased to 100% when the recommended bacterial inoculum was doubled and the test was read strictly after 120 min of incubation. The Rapidec Carba NP test is a useful tool for the reliable confirmation of carbapenemase-producing Enterobacteriaceae isolates. The test should be read strictly after 120 min of incubation and the inoculum should be larger than recommended by the manufacturer.     

Simplified Protocol for Carba NP Test for Enhanced Detection of Carbapenemase Producers Directly from Bacterial Cultures

We compared carbapenemase detection among 266 Gram-negative bacilli (161 carbapenemase producers) using the Carba NP tests issued by the CLSI (CNPt-CLSI) and a novel protocol (CNPt-direct) designed for carbapenemase detection direct from bacterial cultures (instead of bacterial extracts required by the CLSI tests). The specificities were comparable (100%), but the CNPt-direct was more sensitive (98% versus 84%). The CNPt-direct was easier to perform due to the direct use of colonies and offered a more robust detection of carbapenemase producers.     

The prediction values of carbapenemase detection methods and carbapenem susceptibility testing for clinical outcomes of patients with Acinetobacter bacteremia under carbapenem treatment

BackgroundCarbapenem-resistant Acinetobacter species have emerged as notorious pathogens causing nosocomial infections. Several phenotypic methods have been developed for detecting carbapenemase production in Enterobacteriaceae. The accuracy of these methods in the prediction of carbapenemase production in Acinetobacter species has not been studied well.MethodsThis retrospective study enrolled adult patients with Acinetobacter bacteremia from four medical centers in Taiwan between 2012 and 2016. Their demographics and clinical outcomes were recorded. The carbapenem susceptibility of the Acinetobacter species was determined using the agar diffusion method. The carbapenemase genes were detected by PCR. Four phenotypic methods, including the modified Hodge test (MHT), modified carbapenem inactivation method (mCIM), Carba NP test, and CarbAcineto NP test were carried out to determine the production of carbapenemase.ResultsWe analyzed 257 adults who received initial carbapenem monotherapy for the treatment of Acinetobacter bacteremia. Shock within three days of bacteremia and acquisition of carbapenem non-susceptible isolates were independently associated with a higher 14-day and 30-day mortality in patients with Acinetobacter bacteremia. Among the four phenotypic tests for carbapenemase detection, MHT using the imipenem disc displayed the greatest sensitivity (94%; 95% confidence interval [CI], 89–97%) and specificity (81%; 95% CI, 73–88%) for predicting imipenem non-susceptibility.ConclusionCarbapenem non-susceptibility and shock were independent risk factors for mortality in patients with Acinetobacter bacteremia. The MHT could predict the carbapenem susceptibility of Acinetobacter isolates. It is a cheap and quick assay, which could be applied in clinical practice.     

Comparative Study of a Novel Biochemical Assay,the Rapidec Carba NP Test,for Detecting Carbapenemase-Producing Enterobacteriaceae

The novel biochemical test, the Rapidec Carba NP (RCNP), was evaluated using carbapenemase- and non-carbapenemase-producing Enterobacteriaceae isolates. The RCNP test was compared with the Carba NP test (CNP) and the modified Hodge test. Compared to the CNP test, the RCNP test had identical sensitivity (96%) and lower specificity (93% versus 100%). The medium used to culture the isolates significantly affected test sensitivity and specificity. The RCNP test was quicker and easier to perform than the other tests.     

Comparison of MALDI-ToF MS with the Rapidec Carba NP test for the detection of carbapenemase-producing Enterobacteriaceae

Although carbapenemase-producing Enterobacteriaceae (CPE) have become a serious public health issue, their detection remains challenging. The aim of this study was to implement a test based on imipenem hydrolysis by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-ToF MS), using 65 strains producing or not a carbapenemase. Then, we compared its performance to that of the Rapidec Carba NP test using 20 additional strains. The MS-based test effectively discriminated between CPE and other non-carbapenem-susceptible strains compared to the Rapidec Carba NP test (sensitivity 100% and 92%, specificity 94% and 92%, respectively). The MS-based test gave less difficulty in interpretation than the colorimetric Rapidec Carba NP test. MALDI-ToF gave a result in less than one hour and limited the use of expensive molecular assays. In conclusion, the hydrolysis test based on MALDI-ToF MS can detect clinically relevant CPE isolates in routine practice. This technology, also described to screen for carbapenem resistance in Pseudomonas aeruginosa and Acinetobacter baumannii complex strains, also seems to be interesting in routine practice for these pathogens.     

Detection of carbapenemases in Enterobacteriaceae: a challenge for diagnostic microbiological laboratories

Carbapenemase-producing bacteria have now spread all over the world. Infections caused by those bacteria are difficult to treat. Therefore, there is an urgent need for accurate and fast detection of carbapenemases in diagnostic laboratories. In this review, we summarize screening methods for suspected isolates, direct assays for confirmation of carbapenemase activity (e.g. the Carba NP test and matrix-assisted laser desorption ionization time-of-flight mass spectrometry carbapenem hydrolysis assay), inhibitor-based methods for carbapenemase classification, and molecular-genetic techniques for precise identification of carbapenemase genes. We also propose a workflow for carbapenemase identification in diagnostic laboratories.     

Rapidec Carba NP Test for Rapid Detection of Carbapenemase Producers

Performances of the Rapidec Carba NP test (bioMérieux) were evaluated for detection of all types of carbapenemases in Enterobacteriaceae, Acinetobacter baumannii, and Pseudomonas aeruginosa. In less than 2 h after sample preparation, it showed a sensitivity and specificity of 96%. This ready-to-use test is well adapted to the daily need for detection of carbapenemase producers in any laboratory worldwide.     

CarbAcineto NP Test for Rapid Detection of Carbapenemase-Producing Acinetobacter spp.

Multidrug-resistant Acinetobacter baumannii isolates, particularly those that produce carbapenemases, are increasingly reported worldwide. The biochemically based Carba NP test, extensively validated for the detection of carbapenemase producers among Enterobacteriaceae and Pseudomonas spp., has been modified to detect carbapenemase production in Acinetobacter spp. A collection of 151 carbapenemase-producing and 69 non-carbapenemase-producing Acinetobacter spp. were tested using the Carba NP test and a modified Carba NP protocol (the CarbAcineto NP test) in this study. The CarbAcineto NP test requires modified lysis conditions and an increased bacterial inoculum compared to those of the original Carba NP test. The Carba NP test detects metallo-β-lactamase producers but failed to detect the production of other carbapenemase types among Acinetobacter spp. In contrast, the newly designed CarbAcineto NP test, which is rapid and reproducible, detects all types of carbapenemases with a sensitivity of 94.7% and a specificity of 100%. This cost-effective technique offers a reliable and affordable technique for identifying carbapenemase production in Acinetobacter spp., which is a marker of multidrug resistance in those species. Its use will facilitate the recognition of these carbapenemases and prevent their spread.