目的 比较干化学法、尿总蛋白/尿肌酐比值(PCR)和尿清蛋白/尿肌酐比值(ACR)3种尿蛋白筛查方法的一致性,并分析其在健康体检中的应用价值.方法 选取2020年8月11-30日解放军总医院第六医学中心健康管理科936例健康体检者晨尿标本,同时进行干化学法尿蛋白定性/半定量测定,比色法或比浊法尿总蛋白、尿清蛋白和尿肌酐定量测定,研究干化学法、PCR和ACR 3种检测方法阴阳性符合率,干化学法少量蛋白尿与ACR微量清蛋白尿测定的一致性,以及尿蛋白阳性体检者标本情况等.结果 当ACR<30 mg/g时,PCR符合率97.6％,干化学法符合率98.9％,阴性符合率高;当ACR在30～300 mg/g时,PCR假阴性率为28.7％,干化学法假阴性率为78.3％,阳性符合率低;当ACR>300 mg/g时,PCR符合率为100.0％,干化学法符合率84.6％,其中2例干化学法假阴性均因高尿酸血症口服碳酸氢钠所致,阳性符合率高.36例干化学法1+(半定量250～<750 mg/L)体检者标本中,对应尿清蛋白水平仅10例在此区间内,符合率为27.8％,这与研究人群年龄和代谢状态相关.118例3种检测方法结果不一致体检者标本中,8例干化学法阳性、PCR和ACR均阴性,临床特征包括3例糖尿病和4例高血压/高血脂患者.155例尿蛋白阳性标本中,干化学法尿蛋白假阴性率68.1％,假阳性率0.1％;ACR假阴性率16.2％,假阳性率1.8％;PCR假阴性率20.6％,假阳性率0.8％.结论 尿蛋白检测应针对不同对象选择不同检测和评价方法,尿蛋白结果的解读应纳入患者近期饮食、口服药物和代谢状态.

目的 建立基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)霉菌数据库,提高引起临床相关感染的各种霉菌的鉴定能力.方法 收集临床标本来源的霉菌266株,纯培养后进行霉菌18S rDN A扩增测序,获得菌种鉴定结果.选取210株霉菌用于数据库的构建,通过甲酸萃取法提取蛋白,应用M A LDI-T O F MS配套的FlexControl软件收集蛋白谱图,采用Biotyper软件构建评价数据库.其余56株霉菌用于数据库鉴定能力的验证,分别用商品化数据库、自建的MALDI-TOF MS数据库和扩展数据库进行菌种鉴定,应用 χ2检验比较数据库的鉴定率差异.结果 自建本地化MALDI-TOF MS霉菌鉴定数据库包含19个菌属,67个菌种,共计204株霉菌的质谱图.用于数据库验证的56株霉菌中,用商品化数据库鉴定正确18株,鉴定正确率为32.1％,用扩展数据库鉴定正确51株,鉴定正确率为91.1％,差异有统计学意义(P<0.05).结论 自建霉菌数据库提高了霉菌的鉴定能力,从而有助于临床霉菌感染的病原诊断和治疗.

目的 探讨红细胞平均体积(MCV)对XN-2000血液分析仪电阻抗法(PLT-I)和荧光染色法(PLT-F)计数患者血小板结果的影响.方法 采用Sysmex XN-2000血液分析仪对286例患者抗凝血分别用PLT-I和PLT-F进行血小板计数,计数结果根据MCV大小分为3组,A组:MCV<72 fL,B组:72 fL≤MCV≤82 fL,C组:MCV>82 fL;将两种方法检测的血小板结果进行统计学分析.结果 A组PLT-I和PLT-F两种方法计数结果差异有统计学意义(P<0.05),B组和C组这两种方法计数结果差异均无统计学意义(P<0.05).结论 临床进行血常规血小板计数检测时,MCV越大,PLT-I和PLT-F二者计数结果差异越不明显,MCV<72 fL时,PLT-I和PLT-F血小板计数结果有明显差异,提示MCV<72 fL时采用PLT-F对血小板计数结果进行复检,可有效避免小红细胞导致的血小板计数假性增高.

目的 探讨围生期孕妇无乳链球菌(GBS)的临床感染情况及耐药性特征,为临床合理使用抗菌药物提供参考.方法 对2017年4月至2020年3月在该院产检的围生期孕妇6313例进行GBS检测,根据检测结果 分为GBS阳性组和GBS阴性组,比较2组孕妇的一般资料及妊娠结局,分析GBS对10种抗菌药物的耐药率.结果 6313例围生期孕妇中,GBS感染率为7.16％(452/6313);2组孕妇年龄,妊娠期糖尿病、妊娠期高黏稠血症、妊娠期阴道炎检出率比较,差异无统计学意义(P>0.05),2组双胎妊娠、不良孕史比较,差异有统计学意义(P<0.05);2组在早产、羊水污染、产后出血、剖宫产率方面比较,差异无统计学意义(P>0.05),2组胎儿窘迫、先兆流产、胎膜早破发生率比较,差异有统计学意义(P<0.05).分离出的452株GBS对利奈唑胺、青霉素、奎奴普丁/达福普汀、替加环素、万古霉素、替考拉宁、氨苄西林敏感率为100.00％,对克林霉素、左氧氟沙星、四环素的敏感率分别为28.98％、44.91％、67.92％.结论 围生期孕妇GBS感染易造成胎儿窘迫、先兆流产及胎膜早破等不良妊娠结局,临床上应加强围生期孕妇GBS感染的防控工作,目前青霉素仍是治疗GBS的首选药物.

目的 探讨HB&L微生物培养仪对尿路感染(UTI)的诊断价值.方法 收集2019年6-12月在该院就诊的疑似UTI的277例患者作为研究对象,采集患者尿液标本,分别采用尿培养及HB&L微生物培养仪判断患者UTI情况,采用质谱仪鉴定阳性患者菌种分布情况,分析HB&L微生物培养仪培养对UTI的诊断价值.结果 HB&L微生物培养仪培养时间与尿培养时间比较,差异有统计学意义(P<0.05);277例尿液标本中,尿培养阳性检出率(23.47％)与HB&L微生物培养仪阳性检出率(23.10％)比较,差异无统计学意义(χ2=0.010,P>0.05);以尿培养结果作为金标准,HB&L微生物培养仪的灵敏度、特异度、准确度、阳性预测值、阴性预测值分别为96.92％、99.53％、99.28％、98.44％、99.06％;单一菌感染55例,混合菌感染10例,采用HB&L微生物培养仪鉴定菌种检出率为90.67％,错误率为1.33％,未检出率为8.00％.结论 与尿培养比较,HB&L微生物培养仪能够有效缩短培养时间,且二者阳性检出率比较无差异,HB&L微生物培养仪鉴定菌种的检出率较高.

目的 评价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)用于临床常见酵母样真菌鉴定的截断值,为酵母样真菌鉴定提供适宜的评分标准.方法 收集从临床各类标本中分离出的酵母样真菌494株,将纯培养菌株直接涂抹至靶板上,滴加甲酸和基质液后进行MALDI-TOF MS鉴定,记录鉴定分值;同时进行基于真菌18srDNA的测序验证,以测序为金标准,评价MALDI-TOF MS鉴定的准确性;应用SPSS软件绘制ROC曲线,得出MALDI-TOF MS截断值,应用 χ2检验比较本次研究得出的截断值与机器推荐截断值鉴定的正确率和错误率.结果 27个菌种,463株酵母菌质谱鉴定与测序结果一致,鉴定正确率为93.7％;截断值为1.6时ROC曲线下面积最大,对应的敏感度为0.86,特异性为0.88;经 χ2检验,与原截断值2.0相比,截断值1.6可将正确鉴定率提高至77.7％,差异有统计学意义(P<0.001).结论 根据前处理方法的不同计算酵母样真菌鉴定适宜的截断值,将有助于提高MALTI-TOF MS酵母样真菌的鉴定能力.

目的 探讨荧光染色法在真菌镜检中的应用价值.方法 收集350例不同来源的临床标本,直接涂片后分别进行革兰染色和荧光染色镜检,同时接种沙保罗培养基进行真菌培养鉴定,3种方法任意一种为阳性,则认为该标本为真菌检测阳性,并以此为标准来计算3种方法的检出率、灵敏度和阴性预测值,应用χ2检验评价3种方法的差异.结果 350例临床标本应用荧光染色法、革兰染色法和真菌培养鉴定法的检出率分别为53.4％、11.4％和43.4％,灵敏度分别为86.6％、18.5％和70.4％,阴性预测值分别为82.2％,43.2％和67.7％,各组数据比较,差异均有统计学意义(P<0.05).结论 荧光染色法在真菌检测中显示出一定的优势,值得推广应用.

目的 建立海洋细菌基质辅助激光解吸电离飞行时间质谱鉴定数据库,为海洋细菌引起各类感染的病原快速鉴定提供支持.方法 本中心课题组2006-2016年收集国内外6个海域158个采样点海洋细菌530株,分离培养后进行细菌16srDNA扩增测序,应用FlexControl软件收集蛋白谱图,Biotyper软件构建评价数据库.分别用仪器商品化数据库、自建数据库和扩展数据库进行菌种鉴定,比较各数据库的鉴定差异.结果 自建本地化海洋细菌鉴定数据库包含32个菌属,82个菌种,共计421株海洋细菌谱图.以16srDNA测序鉴定结果 为金标准,验证菌株用商品化数据库鉴定时正确80株,鉴定正确率80.8％,用扩展数据库鉴定时正确97株,鉴定正确率98.0％,差异有统计学意义(P<0.001).结论 自建海洋细菌鉴定数据库菌株的鉴定能力较商品化数据库更强,有助于海洋细菌引起相关感染的诊断和治疗.

目的 通过对实验室自动化系统运行数据进行分析,调整系统运行参数,改进工作流程,缩短标本周转时间.方法 对3631份标本运行数据进行统计分析,计算标本周转时间,并进行超时原因分析.针对超时最严重的肿瘤标志物检测标本,改变检测流程,对流程改变前后标本周转时间进行比较.对人工干预标本按照原因进行分类,并针对问题进行改进,对改进前后人工干预标本数量及占比进行分析.结果 标本周转时间超过4 h的标本共313份,占总标本数的8.62％.其中因标本交接超时的标本在超时标本总数中占比最大,为66.77％.肿瘤标志物标本检测流程改进后标本周转时间明显缩短,这类标本改进前占超时标本总数的52.47％,改进后为24.76％.条码异常、血量不足、离心问题3种原因造成的人工干预标本占总数的90.00％以上,针对3种主要原因,实验室采取对应措施后,人工干预标本占比下降至4.21％.结论 实验室通过自动化系统运行数据分析,可针对性地解决瓶颈问题,提高工作效率.

目的 运用基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)联合分离胶法快速鉴定血培养阳性标本,探讨快速鉴定法的符合率.方法 运用分离胶法提取阳性血培养瓶中富集的菌落,应用MALDI-TOF-MS对经前处理的血培养阳性标本进行直接鉴定,鉴定结果与传统的VITEK-2 Compact全自动细菌分析仪鉴定结果进行比较,若有不符,以16SrRNA序列分析结果作为金标准.结果 在实验的200株阳性血培养标本中,革兰阳性菌的鉴定符合率达79.5％;革兰阴性菌鉴定符合率达85.7％;真菌的鉴定符合率达20.0％.由此可见,革兰阴性菌的鉴定符合率高于革兰阳性菌,二者鉴定符合率均远高于真菌.两种及两种以上细菌混合感染的阳性标本直接鉴定还存在一定困难,其直接鉴定方式还需进一步探讨.应用两种鉴定方法得出不同结果的12株阳性标本,16SrRNA序列分析结果均与VITEK-2 Compact全自动细菌分析仪鉴定结果一致.结论 MALDI-TOF-MS联合分离胶促凝管直接检测血培养阳性标本,对血流感染中主要病原菌的鉴定符合率较高,且迅速简便,此方法可将细菌更快地鉴定到种属,使临床能够进行早期治疗,有效控制菌血症的病死率和院内感染.

本文阐述了本院基于试剂消耗品的管理平台的建设与实践,以提高实验室试剂消耗品的管理水平.以规范化管理为导向,以高效服务为宗旨,以信息技术手段为依托,建设信息化、全面化管理平台.试剂耗材管理平台的建立加强了试剂耗材管理的信息化、规范化及专业化,建立了科学完善的管理体制,提升了服务保障功能.

目的 运用MALDI-TOF-MS技术建立鲍氏志贺菌的蛋白指纹图谱并将其添加至数据库,实现对临床细菌的快速鉴定,便于临床医生对疾病进行快速而有效的控制.方法 采用甲酸萃取法进行鲍氏志贺菌蛋白指纹图谱的添加,研究不同的标本前处理方法对鉴定结果的影响,并用新分离的鲍氏志贺菌和大肠埃希菌对图谱进行验证.结果 采用新的图谱鲍氏志贺菌提取法鉴定准确率为95％,直接涂抹法准确率为79％,对大肠埃希菌的鉴定准确率为97％.鲍氏志贺菌不同处理方法的鉴定准确率比较,差异有统计学意义(P<(0.05).结论 新添加的鲍氏志贺菌蛋白指纹图谱能快速准确地鉴定鲍氏志贺菌,此图谱可以应用于鲍氏志贺菌的临床快速诊断.

Objective To establish a method for the identification of Vibrio parahaemolyticus based on Taqman-fluorescence probe quantitative PCR method targeting toxR gene.Methods Taking the standard strain of Vibrio parahaemolyticus (VPJS421) and other ommon pathogenic bacteria'standard strain as the research object,using the bio-software to design specific PCR primers and Taqman probe of Vibrio parahaemolyticus toxR gene and detected by fluorescence quantitative PCR instrument.Results ①The designed primers could amplify specific bands.②The amplification efficiency of the 0.5 μl probe in the amplification system was better than that of the 1.0μl probe.③The detection sensitivity of toxR gene of Vibrio parahaemolyticus by Taqman fluorescence quantitative PCR was 10-1 mg/L.④The detection method did not show positive amplification in detection of Enterococcus f aecalis,Staphylococcus aureus,Saprophytic staphylococcus,Enterobacter hormaechei,Pseudomonas aeruginosa,Escheri ch ia coli,Vibrio al ginol yticus,Vibrio vulnficus,Vibrio metschnikovii and Wbrio furnissii 10 other common pathogenic bacteria.The specificity was 100％.Conlusion The fluorescence quantitative PCR method for the identification of Vibrio parahaemolyticus was successfully established.The method was sensitivty and specificity,and it is suitable for rapid detection of Wbrio parahaemolyticus and has a good application value.

目的 通过对180株金黄色葡萄球菌临床分离株MecA、Nuc基因的检测,以期选择出一种适合临床的并能简便、快速、准确检测耐甲氧西林金黄色葡萄球菌(methicillin resistant Staphylococcus aureus,MRSA)的方法.方法 细菌鉴定及药敏采用全自动细菌鉴定和药敏分析仪.MecA、Nuc基因检测采用荧光定量聚合酶链反应(fluorescence quantitative polymerase chain reaction,FQ-PCR)法.结果 FQ-PCR对180株金黄色葡萄球菌MecA、Nuc基因检出率为75.0%,全自动细菌鉴定和药敏分析仪对180株金黄色葡萄球菌的MRSA检出率为72.8%,2种检测方法比较差异无统计学意义(P>0.05).全自动细菌鉴定和药敏分析仪对MRSA的检测正确率为94.8%.结论 FQ-PCR检测MRSA正确率高,只需要1~2 h即可完成.该方法快速、简便、准确,值得推广应用.

The aim of this study was to develop a rapid detection assay to identify methicillin-resistant Staphylococcus aureus by simultaneous testing for the mecA, nuc, and femB genes using the loop-mediated isothermal amplification (LAMP) method. LAMP primers were designed using online bio-software (http://primerexplorer.jp/e/), and amplification reactions were performed in an isothermal temperature bath. The products were then examined using 2% agarose gel electrophoresis. MecA, nuc, and femB were confirmed by triplex TaqMan real-time PCR. For better naked-eye inspection of the reaction result, hydroxy naphthol blue (HNB) was added to the amplification system. Within 60 min, LAMP successfully amplified the genes of interest under isothermal conditions at 63 °C. The results of 2% gel electrophoresis indicated that when the Mg2+ concentration in the reaction system was 6 μmol, the amplification of the mecA gene was relatively good, while the amplification of the nuc and femB genes was better at an Mg2+ concentration of 8 μmol. Obvious color differences were observed by adding 1 μL (3.75 mM) of HNB into 25 μL reaction system. The LAMP assay was applied to 128 isolates cases of methicillin-resistant Staphylococcus aureus, which were separated from the daily specimens and identified by Vitek microbial identification instruments. The results were identical for both LAMP and PCR. LAMP offers an alternative detection assay for mecA, nuc, and femB and is faster than other methods.

Objective Two identification systems,Matrix assisted laser desorption ionization time of flight mass spectrometry system and VITEK 2 Compact were compared for the efficiency of identification of clinically relevant bacteria.Methods The 558 selected clinical isolates tested in this study were isolated from clinical samples of blood,urine,cerebrospinal fluid,secretion,sputum et al,and were obtained from the navy general hospital from Aug 2015 to Sep 2015,then the isolated strains were identification with the use of Bruker Biotyper bacterial identification system and VITEK 2 Compact bacterial identification system at the same time,at last the identification result identified by the two devices were compared.Results the 558 strains bacteria from clinical samples were divided into 28 genuses by Bruker Biotyper and were divided into 24 genuses by VITEK 2 Compact.There are 22 genera that were identified by both two devices,among the result of two devices,the agreement rate of identification of species reached 91.0％,the agreement rate of identification of genus reached 94.8％.While,the salmonella which take a rate of 0.2％ in clinical isolates are identified to genus by both two instrument.The agreement rate of identification of species of gram-negative bacteria was not significantly more than gram-positive bacteria and fungus(P ＞0.05).Conclusion The Bruker Biotyper system is rapid,accurate,and convenient,low cost in the identification and diagnosis in the clinical laboratory.Still,the database needs to be further improved.

Objective To investigate the changes of drug resistance and class Ⅰ integron in clinically isolated Pseudomonas aeruginosa(PA) in our hospital form January 2013 to December 2015.Methods Clinically isolated PA strains were divided into the 3 time periods of 2013,2014 and 2015.Their resistance to 22 commonly used antibacterial drugs was investigated by adopting the VITEK-2;200 strains were randomly selected from the isolated strains during these 3 time periods.Class Ⅰ integron was detect by PCR.Results The detected PA in these 3 time periods had 366,437 and 520 strains respectively.The drug resistance of Pseudomonas aeruginosa to 22 commonly used antibacterial drugs was remarkably increased(P<0.05),especially in ICU.The detection rate of class Ⅰ integron positive bacteria was gradually increased year by year,moreover the drug resistance rate of class Ⅰ integron positive bacteria was significantly higher than that of class Ⅰ integron negative bacteria (P<0.01).Conclusion The drug resistance rate of PA in this hospital is higher.The proportions of multi-drug resistance and classⅠ integron are significantly increased.The hospital infection detection and drug-resistant bacterial monitoring should be strengthened to further standardize the use of antibacterial drugs.

Objective To explore the application of LIS management group in hospital software and hardware management.Methods The objective,responsibilities and management procedure of hospital LIS management group were introduced,and its effects were studied in hospital software and hardware management.Results Hospital LIS management group optimized LIS management procedure,decreased fault rate and shortened the time for fault handling.Conclusion LIS management group promotes hospital digitalization and clinical laboratory department management.

Tuberculosis is caused by mycobacterium, a potentially fatal infectious bacterium. In recent years, TB cases increased in the whole world. WHO statistics data shows that the world's annual tuberculosis incidence was 8~10 million with about 3 million deaths. Several studies have shown that susceptibility to tuberculosis may be associated with IFNGR1 gene polymorphisms. Here, we report the distribution frequency of IFNGR1 gene polymorphisms in 103 cases of IGA-negative patients and 100 cases of IGA-positive patients from China by sequencing the IFNGR1 proximal ~750 bp promoter region. We found a total of 5 types of site mutations: -611 (G/A), -56 (T/C), -255 (C/T), -359 (T/C), and -72 (C/T). The two main types of gene polymorphisms among the IGA-negative and IGA-positive groups were -611 (G/A), with mutation rates of 88.3% and 78.4%, respectively, and -56 (T/C), with mutation rates of 84.5% and 83.8%, respectively, which had no statistical significance, and there was no correlation with the incidence of tuberculosis.

Objective To probe into the superiority of MALDI-TOF MS in the identification of clinical yeast to provide the basis for early diagnosis and treatment of clinical fungal infection.Methods A total of 206 strains of yeast clinically isolated in our hospital from August 2015 to March 2016 were collected and identified with MALDI-TOF mass spectrometer,VITEK2 compact microbial analyzer and chromogenic culture respectively.The ITS sequencing results were used as the final confirmation one.The accuracies of identifying yeast were compared among these 3 kinds of method.Results The accuracy rates of 3 kinds of method for identifying yeast were 82.5% for VITEK2 Compact,67.0% for chromogenic method and 92.7% for MALDI-TOF mass spectrometry.The difference between MALDI-TOF and VITEK2 compact was statistically significant(P=0.006),and the difference between MALDI-TOF and chromogenic method was statistically significant(P=0.001).Conclusion Compared with the VITEK2 Compact microbial analyzer and the color developing method,the MALDI-TOF mass spectrometer has the advantages of rapidness,simple operation,accurate results and so on.