Laboratory evaluation of the CHROMagar KPC medium for identification of carbapenem-nonsusceptible Enterobacteriaceae

CHROMagar KPC was evaluated with 53 carbapenem-nonsusceptible strains. KPC-producing Klebsiella pneumoniae and Enterobacter cloacae grew efficiently and yielded metallic blue colonies (100% sensitivity). Poor or no growth or atypical colony color were evident with KPC-producing Escherichia coli (71.4% sensitivity) and ertapenem-resistant KPC-nonproducing K. pneumoniae (72.7% sensitivity). CHROMagar KPC appears applicable for processing carbapenem-resistant Enterobacteriaceae surveillance specimens, but its performance might possibly be suboptimal in the presence of KPC-negative ertapenem-resistant strains or for detection of KPC-producing E. coli.     

定位显色培养基在细菌快速初步鉴定中的应用

目的评价定位显色培养基在细菌快速初步鉴定中的作用。方法将3313株临床菌株和145株室间质评菌株点种于定位显色培养基上,观察显色特点,总结细菌显色规律。结果不同种属的细菌在定位显色培养基上具有一定的显色规律。结论定位显色培养基对细菌的快速初步推测和鉴定具有重要的应用价值,具有成本低廉、有助于提高工作效率等优势,值得推广应用。     

简略快速法在大肠埃希菌鉴定中的应用价值研究

目的比较简略快速法与自动生化鉴定法在大肠埃希菌鉴定中的能力差异。方法使用快速简略鉴定法和自动生化鉴定法对临床标本分离的疑似大肠埃希菌进行鉴定,比较两种方法结果的一致性及检测时间。结果 492株疑似大肠埃希菌经快速鉴定法检出248株大肠埃希菌及244株非大肠埃希菌;该248株大肠埃希菌中,有231株被自动生化鉴定仪确认为大肠埃希菌,而244株非大肠埃希菌中,有7株被确认为大肠埃希菌。简略快速法阳性预测值及阴性预测值分别为93.1%(231/248)及97.1%(237/244)。使用自动生化鉴定法耗时2.5~7.0h,平均(4.12±1.08)h;使用简略快速法耗时0.5~2.0h,平均(1.08±0.45)h,差异具有统计学意义(t=-40.252,P0.001)。结论使用简略快速法鉴定疑似大肠埃希菌可以获得接近于自动生化鉴定法的结果,且更加节省时间。     

血浆替代血清行常规生化检测缩短正流转时间的可行性探讨

目的探讨肝素锂抗凝血浆替代血清进行常规生化检测缩短正流转时间(TAT)的可行性。方法应用日立H7600-20全自动生化分析仪对抽取的110例患者肝素锂抗凝血浆和血清中的35项生化检测结果进行比对分析。结果在肝素锂抗凝血浆与血清的35项生化检测结果中,清蛋白(ALB)、γ-谷氨酰基转移酶(GGT)、总胆汁酸(TBA)、总胆红素(TB)、氯(Cl)、钙(Ca)、镁(Mg)、肌酐(CRE)、尿素(BUN)、尿酸(UA)、胱抑素C(CysC)、β2-微球蛋白(β2-MG)、肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)和高密度脂蛋白胆固醇(HDL-C)共16项指标的检测结果比较差异无统计学意义(P0.05);而钠离子(Na+)、钾离子(K+)、二氧化碳(CO2)、磷(P)、铁(Fe)、丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、总蛋白(TP)、前清蛋白(PreA)、α-L-岩藻糖苷酶(AFU)、三酰甘油(TG)、总胆固醇(TCH)、低密度脂蛋白胆固醇(LDL-C)、载脂蛋白A1(ApoA1)、载脂蛋白B(ApoB)、脂蛋白a[Lp(a)]、血糖(GLU)、直接胆红素(DB)共19项指标的检测结果比较差异有统计学意义(P0.05)。结论肝素锂抗凝血浆与血清中常规生化检测结果不完全一致,肝素锂抗凝血浆可选择性地替代血清进行部分急诊生化检测以缩短TAT。     

3-氨基苯硼酸联合乙二胺四乙酸碳青霉烯酶抑制增强试验检测肠杆菌目细菌产碳青霉烯酶的研究

目的 了解3-氨基苯硼酸(PBA)联合乙二胺四乙酸(EDTA)碳青霉烯酶抑制剂增强试验(PBA-EDTA法)用于检测肠杆菌目细菌产碳青霉烯酶的检测结果.方法 使用PBA-EDTA法测定275株经金标准PCR及DNA测序已明确为产碳青霉烯酶肠杆菌目细菌产碳青霉烯酶结果的符合率,并与CLSI推荐的mCIM联合eCIM改良碳...     

常规生化检查在初诊多发性骨髓瘤患者中的应用价值

目的 探讨常规生化指标在多发性骨髓瘤(MM)中的应用价值.方法 回顾性分析天津市第三中心医院2004年1月至2010年5月收住院的63例初诊MM患者的临床资料,根据同时检测的血清清蛋白水平对血钙进行校正,然后对MM患者进行Durie-Salmon(DS)分期,分析常规生化指标与MM分期的关系.结果 经血清钙校正后Ⅰ、Ⅱ、Ⅲ期MM患者分别为15例、9例和39例,各期之间球蛋白、清蛋白、β2-微球蛋白及血沉的差异均具有统计学意义(P<0.05),而乳酸脱氢酶和C反应蛋白的差异没有统计学意义(P>0.05).β2-微球蛋白与C反应蛋白呈正相关,与清蛋白、乳酸脱氢酶、血沉则无明显相关(P>0.05).结论 球蛋白、清蛋白和β2-微球蛋白的浓度在MM不同分期之间呈梯度分布,可以为MM分期提供辅助信息,反映MM的病情严重程度.     

循证检验医学在中国驻黎巴嫩维和二级医院生化检验质量控制中的应用

目的利用循证检验医学的观点寻求一种在维和行动中控制生化检验质量的操作办法。方法通过查询有关国际维和生化检验质量控制方面的文献报道,进行比较鉴别,结合5年多的维和检验工作经验,摸索出一套可行的方法,并进行了一个任务期的实践评价。结果在质量控制体系的监控下,向临床提供准确、可靠的检验报告,制订和实施的检验生化质量控制方法经济、实用、可靠,能保障维和医疗检验生化报告的有效性。结论循证检验医学对维和医疗机构生化检验的质量控制具有重要意义。     

Effect of hyperthyroidism and propylthiouracil treatment on liver biochemical tests

Liver biochemical test (LBT) changes can be commonly observed in hyperthyroid patients. Those kinds of changes could also be observed because of propylthiouracil (PTU) therapy. We prospectively evaluated LBT changes because of PTU use for 1 year in patients who had been diagnosed with hyperthyroidism first. We studied 64 patients who had been diagnosed with hyperthyroidism. These patients took at least 1-year PTU treatment. We analysed LBT at diagnosis and after 2 and 12 months of treatment with PTU. Prior to PTU treatment, 30 (46.8%) of the 64 patients had at least one LBT abnormality. We observed at least one LBT abnormality in 11 (32%) patients after 2 months and nine (26%) patients after 12 months of treatment with PTU in 34 patients whose CBT were normal before treatment. We did not observe any deterioration in clinical picture and bilirubin levels. Elevated serum LBT during the pretreatment and PTU treatment period is common and usually transient and asymptomatic. PTU could be used for long times by lowering the dose and close follow-up in patients who have elevated LBT during the pretreatment and after PTU treatment period.     

Evaluation of Disk carbapenemase test using improved disks for rapid detection and differentiation of clinical isolates of carbapenemase-producing Enterobacterales

ObjectivesRapid detection of carbapenemase-producing Enterobacterales (CPE) is important to control spread of the resistance. We previously reported that imipenem disks prepared from injectable imipenem-cilastatin could rapidly detect KPC- and NDM-type carbapenemases. In the present study, we evaluated performance of disks of IPM and combined disks of imipenem-tazobactam and imipenem-EDTA, which were prepared from powders of imipenem and inhibitors.MethodsIsolates of Enterobacterales were recovered from specimens of patients at a tertiary care hospital in Korea during January 2017 and March 2018. Routine CPE detection was performed by the CPE surveillance personnel whereas evaluation of the Disk carbapenemase test (DCT) was performed by the other personnel without knowing the results of surveillance. The DCT was carried out by pressing disks on to colonies and rehydrating in Petri plates and observing color change.ResultsThe DCT differentiated 688 of 694 (sensitivity 99.1%) carbapenemase-producing isolates in 2.5–20 min: 630 with KPC, 51 with NDM, three with IMP, one with VIM, two with KPC and IMP, and one with NDM and OXA-181. The DCT failed to detect six OXA- 48-like enzyme-producing isolates, but the modified method using 96-well flat-bottom microplates with mineral oil cover detected all 29 OXA-48-like enzyme-producing isolates in 20–120 min. The DCT was negative for all 440 ertapenem-nonsusceptible, carbapenemase gene-negative isolates (specificity 100%).ConclusionThe procedure of DCT is simple and can differentiate isolates of Enterobacterales with KPC-, NDM-, IMP- and VIM-type carbapenemases rapidly, and the modified DCT can detect isolates with OXA-48-like enzymes rapidly.     

两种显色培养基分离阪崎肠杆菌的应用评价

目的：评价环凯阪崎肠杆菌显色培养基（HKDFIA）和科玛嘉阪崎肠杆菌显色培养基（CMDFIA）的应用效果,为食品中检测阪崎肠杆菌选用培养基提供依据。方法按照 GB/T 4789．40-2010食品卫生微生物学检验阪崎肠杆菌检验,进行阪崎肠杆菌分离及生化鉴定,并分析培养基的分离效果。结果36份样品中,检出阪崎肠杆菌2份,检出率为5．56％;HKDFI 平板和 CMDFI 平板阳性符合率为100％,疑似菌落检出率分别为16．67％和13．89％（P ＞0．05）。结论HKDFIA 和 CMDFIA 用于培养分离阪崎肠杆菌均具有良好的效果,HKDFIA 的性能已达到国外同类产品的水平,可以作为食品中检验阪崎肠杆的选择培养基。     

Prehospital identification of acute coronary ischemia using a troponin t rapid assay

Objective. To evaluate the performance of a rapid assay for cardiac troponin T (cTn-T) in patients with chest pain in the prehospital setting. Methods. A prospective, observational clinical trial in a rural county served by a single emergency medical services system and two emergency departments. Patients fulfilling prehospital criteria to identify acute coronary ischemia (ACI) had a blood sample applied to the cTn-T rapid-assay device. Quantitative analysis of cTn-T was also performed on each sample at a later time. Medical records were reviewed to determine ultimate diagnoses. Non-admitted patients were followed by telephone at one week. Main outcome measures included the sensitivity, specificity, positive predictive value, and negative predictive value of the rapid cTn-T assay for detecting ACI defined as acute myocardial infarction (AMI) or unstable angina (UA) within one week of presentation. Results. Of 87 patients enrolled, 29 were identified with ACI. This included 15 patients diagnosed as having AM1 and 14 patients diagnosed as having UA. The cTn-T rapid-assay device was positive for five of 87 patients (5.7%); three were associated with AMI and two with UA. Measurement of a single cTn-T to detect ACI had a sensitivity of 17.2% (0.058, 0.358), specificity of 100% (0.950, l), positive predictive value of 100% (0.549,1), and negative predictive value of 70.7% (0.609, 0.806). Conclusion. The cTn-T rapid-assay device may be useful in the prehospital setting to identify a small number of patients with ACI. The authors caution, however, that a negative test in the prehospital setting cannot be used to rule out significant disease.     

干化学法和湿化学法对常规急诊生化项目检测的比较

目的和方法 实验观察和比较临床常规急诊项目（Ｋ,Ｎａ,Ｃｌ,ＢＵＮ,ＣＯ２,ＧＬＵ）湿化学法和干化学法的检测。结果：两种方法的各检测项目的线性范围、回收率均符合临床检验要求,重复性良好,批内、批间ＣＶ均在允许范围,ｒ〉０．９８。检验无显著性差异（Ｐ〉０．０５）,呈高度相关。     

Direct microorganism species identification and antimicrobial susceptibility tests from positive blood culture bottles using rapid Sepsityper Kit

IntroductionWe evaluated the performance of Rapid Sepsityper Kit in species identification (ID) and antimicrobial susceptibility testing (AST).MethodsPositive blood culture bottles (n = 227) containing single microorganisms were enrolled. We compared the direct method using Rapid Sepsityper Kit for ID and AST with the conventional method. The analyses of ID and AST were performed using MALDI Biotyper and BD Phoenix platform, respectively.ResultsThe direct ID method correctly identified 89.4% (203/227) of samples, and Gram-negative bacilli (95.2%) had a higher ID rate than Gram-positive cocci (84.4%). Five cases were misidentified, and non-acceptable identification was high among Streptococcus species.Direct AST results were obtained from 168 isolates. Non-acceptable ID occurred among 24 isolates; 4 Streptococcus species, and 31 isolates, which did not grow in the direct AST method, were excluded. A total of 1714 antibiotic susceptibility tests (625 from 69 Gram-positive cocci and 1089 from 99 Gram-negative bacilli) were performed. The direct AST methods showed 98.3% (1685/1714) of categorical agreement (CA), 0.7% (12/1714) of very major errors, 0.2% (4/1714) of major errors, and 0.8% (13/1714) of minor errors. Complete CA was obtained for methicillin-resistant Staphylococcus aureus and extended-spectrum beta-lactamase-producing Escherichia coli.ConclusionsThe direct ID method using Rapid Sepsityper Kit and the direct AST method in combination with the BD Phoenix platform, which was associated with a reduction of turnaround time, may be a reliable approach for blood culture bottles. However, additional validation and further improvements, especially for Gram-positive cocci, would have an impact on microbiological diagnoses.     

Evaluation of LAMP assay using phenotypic tests and PCR for detection of blaKPC gene among clinical samples

BackgroundCarbapenem‐resistant Enterobacteriaceae (CRE) infection constitutes a public health threat, which blaKPC was the major carbapenemases concerned in China. Timely and efficient diagnosis is of paramount importance for controlling the spread of drug‐resistant bacteria. Here, we develop an approach based on loop‐mediated isothermal amplification (LAMP) for rapid confirmation of blaKPC within 60 min from samples collected.MethodsWe designed primers specific to detect blaKPC and evaluated it for its sensitivity and specificity of detection using real‐time monitoring. Five hundred forty‐six clinical specimens were analyzed by the LAMP assay and compared with the phenotypic tests and PCR. The samples with inconsistent results were further verified by Sanger sequencing.ResultsThe LAMP assay displayed a detection limit of 1 × 10² CFU/ml, which was 10‐fold more sensitive than the PCR. No cross‐reactivity was observed for strains that produced other types of β‐lactamase. Furthermore, we demonstrated concordant results (Kappa > 0.75) between the genotypic method and phenotypic tests for the 546 clinical samples. The data presented in this study suggested that the genotypic method is a reliable assay for identifying blaKPC‐induced CRE in China. The results of the Sanger sequencing indicate that the developed method not only has high accuracy but also meets the need for rapid diagnosis, while the PCR method is prone to false negatives.ConclusionsWe successfully constructed a LAMP technique that can be used for auxiliary diagnosis of CRE, which is faster, cheaper, and more accurate than the PCR. It may therefore be routinely applied for detection of blaKPC producers in routine clinical laboratories.     

日立7600和贝克曼DXC800测量结果可比性的评估

目的使用美国临床和实验室标准协会(CLSI)GP29-A文件方法评估日立7600和贝克曼DXC800测量结果的可比性。方法按生化项目[丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、葡萄糖(GLU)、钾(K)、钠(Na)、氯(Cl)、钙(Ca)、肌酐(Crea)、尿素(Urea)]分别收集单人份血清样本各20份,血清浓度范围均匀覆盖各检验项目的线性范围。每份样本混匀并均分为两份,分别在日立7600全自动生化分析仪和贝克曼DXC800全自动生化分析仪两种不同生化测量系统上进行测量。依据CLSI GP29-A文件方法,以已实现溯源的日立7600全自动生化分析仪为比较测量系统,以贝克曼DXC800全自动生化分析仪为实验测量系统,对两种不同生化测量系统测量结果进行统计并作散点图评估。结果血清生化项目中GLU、K、Na、Cl、Ca、Crea、Urea项目两种测量系统的测量结果在x±D区间内比例在95%以上(不在区间内结果数不超过1个);ALT、AST、TBIL项目两种测量系统的测量结果在x±D区间内比例不足95%(不在区间内结果数大于1个)。结论血清生化项目中GLU、K、Na、Cl、Ca、Crea、Urea两种测量系统的测量结果具有可比性;ALT、AST、TBIL两种测量系统的测量结果不具有可比性。     

基质辅助激光解吸电离飞行时间质谱在临床病原菌鉴定中的应用

目的探讨基质辅助激光解吸电离飞行时间质谱(matrix-assisted laser desorption ionization-time of flight mass spectrometry,MALDI-TOF MS)系统用于快速鉴定临床分离菌的可靠性和实用性。方法收集南京军区南京总医院2013年7~10月自临床标本分离的非重复细菌1 061株,分别使用MALDI-TOF MS和Vitek 2 Compact全自动细菌鉴定仪系统进行鉴定,结果不一致的菌株采用16S r DNA测序验证。结果 1 061株临床分离菌中1 058株(99.7%)经MALDI-TOF MS系统正确鉴定到属水平,1 016株(95.8%)正确鉴定到种,其余3株(0.3%)未给出鉴定结果。5株鉴定结果不一致的菌株经16S r DNA测序确认,结果与MALDI-TOF MS和Vitek 2 Compact鉴定符合率分别为40.0%(2/5)和0(0/5)。结论 MALDI-TOF MS可以作为一个快速、准确和价廉的工具应用于临床分离菌的鉴定。     

COPD患者血气分析和生化试验的结果分析

目的研究各级COPD患者治疗前血气和生化结果的数值、互相的相关性和诊断价值。方法 120例COPD患者进行分级、主要血气和相关生化指标的测定、指标间的相关性分析以及诊断价值研究。结果 PaO2、PaCO2、pH、NA、CL、CO2结合力、TP、ALB、A/G在COPD患者各个分级之间的差异显著（P〈0.05）。ALT、AST、GLB在各个级别之间的差异不显著（P〉0.05）。除了PO2与CO2结合力、PO2与A/G之外（P〉0.05）,其他指标两两之间均具有相关性（P〈0.05）。鉴别COPD患者处于0~Ⅱ级或Ⅲ~Ⅳ级,下列指标的诊断价值中等：PO2、PCO2、CL、CO2结合力。当PCO2/PO2大于0.75、CO2CP/PCO2大于0.60、PCO2/CL大于0.46时,诊断为COPD的Ⅲ级以上的灵敏度和特异度均较高。结论 COPD患者血气和生化指标随着患者的病情进展而变化。PO2与PCO2、PCO2与CO2结合力之间的具有很强的相关性。PO2、PCO2、CL、CO2结合力在其切点下,对患者的病情具有中等水平的鉴别诊断价值。当PCO2小于48.70mmHg时,其灵敏度为100%,即区别0~Ⅱ级与Ⅲ~Ⅳ级COPD的漏诊率为0。PCO2/PO2、CO2CP/PCO2、PCO2/CL的联合指标对COPD患者处于0~Ⅱ级或Ⅲ~Ⅳ级具有优秀的诊断价值。