荧光免疫层析法降钙素原检测试剂盒临床性能评估

目的评估荧光免疫层析法试剂盒检测降钙素原的性能。方法参照美国临床实验室标准化协会(Clinical and Laboratory Standards Institute,CLSI)EP5-A和EP6-A文件方法,对荧光免疫层析法试剂盒(TEBSUN)检测降钙素原的精密度、线性、方法学比对、相对灵敏度和特异性进行评估。结果荧光免疫层析法试剂盒检测降钙素原精密度较好,低浓度和高浓度样本精密度的变异系数分别为8.3%和4.7%;线性验证试验结果显示,在试剂盒标示的检测范围内具有良好的线性梯度关系(r=0.9989);方法学比对结果显示,荧光免疫层析法试剂盒与梅里埃VIDAS酶联免疫荧光法分析系统的降钙素原试剂盒检测结果一致性良好(r=0.9770);在0.5和2.0 ng/ml两个浓度水平,荧光免疫层析法试剂盒相对于酶联免疫荧光法试剂盒的灵敏度和特异性均大于86%,与其测定结果的总体符合率为93.75%。结论荧光免疫层析法试剂盒检测降钙素原的精密度、线性、方法学比对、相对灵敏度和相对特异性等性能评价较好,适用于临床标本检测。     

采用PCR方法快速检测食品中的蛔虫卵

根据蛔虫的特异性基因序列,设计了PCR特异性引物,并进行灵敏性和特异性试验。结果表明:用所建立的PCR检测食品中的蛔虫卵,特异性强,检测灵敏度高,可检测到食品中的一个虫卵。     

食品中金黄色葡萄球菌PCR检测方法的建立

目的:建立快速检测食品中金黄色葡萄球菌的PCR方法。方法:优化金黄色葡萄球菌DNA提取方法并设计基因特异性引物,采用PCR方法扩增金黄色葡萄球菌的耐热核酸酶(nuc)编码基因,验证检测的特异性及灵敏度。结论:所检出的金黄色葡萄球菌在497bp处出现特异片段,该方法具有较高的敏感性和特异性,可作为食品中金黄色葡萄球菌快速检测的手段。     

产肠毒性大肠杆菌DPO-PCR检测方法的建立与应用

双启动寡核苷酸引物(dual-priming oligonucleotide,DPO)是一种新型的引物设计方法,具有设计简易、特异性强以及退火温度范围宽的特点。以产肠毒性大肠杆菌LT基因为靶基因,设计了一对DPO引物,经过对Taq DNA聚合酶、Mg2+、dNTP反应体系因子的优化,建立了产肠毒性大肠杆菌DPO-PCR检测方法,测定了该方法的灵敏度、特异性以及退火温度不敏感性。结果显示,该方法检测灵敏度为1.24×102CFU/ml,在45℃⁶5℃退火温度范围内,均可实现靶基因的高效扩增。该方法特异性强,测试菌株中4株产肠毒性大肠杆菌均为阳性结果,其余菌株为阴性结果,且无非特异性扩增发生。与常规PCR方法相比,DPO-PCR方法不需要对引物参数特别是退火温度反复优化,同时DPO引物的特殊结构又增强了检测特异性,为致病性微生物的快速准确检测提供了新方法。     

彩色多普勒高频超声与血清肿瘤标志物联合诊断小乳腺癌的临床价值

目的:探讨彩色多普勒高频超声与血清肿瘤标志物联合诊断小乳腺癌的临床价值研究。方法:对1200例乳腺彩色多普勒高频超声发现实性肿块,经超声测量直径≤2.0 cm的患者进行血清肿瘤标志物CYFRA21-1、CA15-3及TSGF检查,并追踪手术病理结果,统计两种检查方法联合检验对诊断乳腺癌的灵敏度、特异性及准确率。结果:单独乳腺高频超声检查对诊断乳腺癌的灵敏度87.2%,特异性83.8%,准确率84.5%;单独血清肿瘤标志物对诊断乳腺癌的灵敏度84.1%,特异性81.4%,准确率82.0%,而高频超声和血清肿瘤标志物联合检测的灵敏度、特异性和准确率分别是91.8%、91.5%和90.7%,均高于单独诊断,差异有统计学意义(P<0.05)。结论:彩色多普勒高频超声与血清肿瘤标志物联合诊断小乳腺癌的灵敏度、特异性和准确率较高,具有较高的临床诊断价值。     

红尾皇冠鱼无乳链球菌病LAMP检测方法的建立与应用

以cfb基因保守序列的6个区域设计4条特异性引物,利用链置换DNA聚合酶(Bst DNA polymerase)在恒温(65℃)下保温40 min,建立了无乳链球菌Streptococcus agalactiae的LAMP技术。建立的LAMP方法能够特异性地检测无乳链球菌,检测灵敏度为3.7×101³.7×102cfu/m L,反应前加入显色剂钙黄绿素避免二次污染。现场应用中采用FTA卡采集红尾皇冠鱼Aequidens rivulatus病鱼肝脏组织病原菌,操作简便、快捷。研究表明,针对无乳链球菌cfb基因建立的LAMP检测方法具有较高的特异性及稳定性,尤其是具有快速、简便、成本低等优点,可用于无乳链球菌的野外现场快速检测。     

用于OTA检测的氧化石墨烯/适配体传感器的构建

针对氧化石墨烯(graphene oxide,GO)与适配体之间吸附作用力过强、影响生物传感器检测赭曲霉毒素A(ochratoxin A,OTA)灵敏度的问题,提出了用鲑鱼精子DNA修饰氧化石墨烯表面以减弱其对适配体的吸附作用,提高生物传感器对OTA检测灵敏度的方法。该方法确定了淬灭适配体携带荧光信号的氧化石墨烯最适质量浓度及孵育时间;比较了不同鲑鱼精子DNA浓度对荧光淬灭效率及荧光恢复的影响;对不同浓度的OTA进行检测以确定该生物传感器的检测限及线性检测范围;进行了该生物传感器的特异性分析。研究结果表明,经鲑鱼精子DNA修饰的氧化石墨烯构建的生物传感器对OTA的线性检测范围为20～500nmol/L,检测限为16.7nmol/L,R2=0.99,相对于未经鲑鱼精子DNA修饰的氧化石墨烯构建的生物传感器,其检测灵敏度提高了113倍,而且具有良好的特异性与选择性。     

基于gyrB基因的地衣芽孢杆菌PCR快速检测方法的建立

根据地衣芽孢杆菌的gyrB基因序列设计一对引物,扩增片段大小为507bp的基因片段,该方法对地衣芽孢杆菌DNA的扩增结果为阳性,对照菌株扩增结果均为阴性,对地衣芽孢杆菌检测的灵敏性为1pg总DNA量,成功建立了地衣芽孢杆菌PCR检测方法,该方法具有快速、灵敏度高、特异性强等优点,为地衣芽孢杆菌的分离及鉴定奠定了良好的基础。     

副猪嗜血杆菌环介导等温扩增快速检测方法的建立

为了建立一种简便、快速、灵敏、特异的副猪嗜血杆菌(Hps)检测方法,以Hps的16S rRNA保守区域片段为靶序列,选择6个特异性区域,设计4条特异性引物F3、B3、FIP和BIP。经优化反应体系、检测灵敏性和特异性,建立Hps的环介导等温扩增(LAMP)快速检测方法。研究结果表明:该方法的最优反应体系是引物c(F3)∶c(FIP)或c(B3)∶c(BIP)为1∶4、Mg~(2+)浓度为2 mmol/L、d NTPs浓度为6 mmol/L,最低检出量为0.241pg/μL DNA。该方法灵敏度是PCR法的100倍,且能够特异性检测出Hps,不能检测出猪链球菌、沙门氏菌、大肠杆菌、枯草芽孢杆菌、胸膜肺炎放线杆菌和金黄色葡萄球菌。     

GBZ-Ⅱ型自动相板测光仪在发射光谱分析中的应用

文章从相板测光仪的几个关键技术入手,对于发射光谱分析方法中Ag、Cu、Pb、Zn、Co、Ni、Sn、Mo等多元素同时测定做了大量实验工作,使得光谱分析中高灵敏度元素Cu与低灵敏度元素Sn、Mo、Ag做到了同板测量,而且光谱法对于多元素测定有其独特优势,不必用化学法分解样品。这样,既节省时间又节约材料,而且方法灵敏度及准确度均达到检测要求。     

“游程总数”法诊断Meta分析中发表偏倚的准确度评价

目的评价利用文献的发表时间信息以＂游程总数＂构建的发表偏倚诊断方法（＂游程总数＂法）的准确度,为其推广应用提供理论依据。方法运用Matlab 7.0软件产生有偏和无偏Meta分析研究模拟数据,计算并比较＂游程总数＂法与Egger法的灵敏度和特异度,对＂游程总数＂法的诊断准确度进行评价。结果 ＂游程总数＂法与Egger法的灵敏度较低（分别为：0.3302±0.0574和0.360 7±0.1856）,而特异度（分别为：0.7062±0.030 8和0.7896±0.0108）较好;＂游程总数＂法的灵敏度和特异度均低于Egger法,但相差幅度小;＂游程总数＂法的灵敏度与Meta分析中的子研究个数有关。结论新构建的＂游程总数＂法与Egger法具有相似的灵敏度和特异度,由于其特殊的理论基础,与基于漏斗图的常用方法联合使用对发表偏倚进行诊断,有利于改善目前灵敏度较低的现状。     

心电图中QT间期和T波面积自动测量的算法

首先研究了ECG中各波特征点的识别,在Q波起点和T波起始点识别的基础上介绍了计算机自动测量QT间期和T波面积的算法;解决了QT间期手工测量时不同测量者测量差异很大以及T波面积无法人工测量的问题;结果表明计算机应用此算法能准确的测量QT间期和T波面积,为临床诊断心肌缺血提供了新的方法,特异度以及敏感度均达到要求.     

唾液及血清c-erbB-2蛋白、CA153检测在乳腺癌诊断中的应用

目的探讨唾液及血清中c-erbB-2蛋白（P185）、CA153检测对乳腺癌的诊断价值。方法采集乳腺癌患者（n=35,乳腺癌组）、乳腺良性疾病患者（n=32,良性组）及健康女性（n=35,对照组）的静脉血及唾液。运用ELISA方法检测唾液及血清中P185水平,电化学发光免疫分析法检测唾液及血清中CA153水平,并计算各项指标的灵敏性、特异性、诊断准确性等。结果乳腺癌组唾液及血清P185、CA153水平均明显高于良性组及对照组,差异有统计学意义（P〈0.05）;良性组与对照组比较,差异无统计学意义（P〉0.05）。唾液标本2项肿瘤标志物检测诊断乳腺癌的敏感性均略低于血清标本,特异性均高于血清标本。唾液及血清标本2项联合检测与单项标志物比较,诊断的敏感性有明显的提高,而且保持了较高的特异性和诊断准确性。结论唾液及血清联合检测c-erbB-2蛋白、CA153均能提高乳腺癌的诊断率,检测唾液中的肿瘤标志物有望在乳腺癌的诊治中得到临床应用。     

某院ICU病房老年侵袭性假丝酵母菌感染患者与血清1,3-β-D-葡聚糖和PCT的关系

目的探讨血清1,3-β-D-葡聚糖及降钙素原（PCT）水平在ICU侵袭性假丝酵母菌感染老年患者中的诊断价值。方法选取47例假丝酵母菌感染老年患者（感染组）及50例老年健康体检者（对照组）,感染组和对照组患者分别于入住ICU、体检时检测1,3-β-D-葡聚糖及PCT水平。结果感染组血清1,3-β-D-葡聚糖及PCT水平均显著高于对照组（P〈0.05）。感染组血清1,3-β-D-葡聚糖诊断受试者工作特征曲线（ROC）曲线下面积（AUC）为0.921;针对老年患者,1,3-β-D-葡聚糖最佳cut-off值为24.2ng·L^-1,诊断敏感度为93.9%,特异性为81.6%;感染组PCT诊断AUC为0.832,针对老年患者,PCT最佳cut-off值为0.55ng·L^-1,诊断敏感度为82.7%,特异性为80.9%;1,3-β-D-葡聚糖诊断效能优于PCT（Z=2.723,P=0.006,P〈0.05）。感染组1,3-β-D-葡聚糖阳性检出率（89.36%）显著高于PCT阳性检出率（44.68%）（χ2=16.000,P〈0.05）。结论血清1,3-β-D-葡聚糖检测对老年患者假丝酵母菌感染诊断具有高度灵敏度与特异性,诊断效能高;PCT检测诊断效能低于1,3-β-D-葡聚糖,但也可辅助临床早期诊断侵袭性假丝酵母菌感染。     

胶体金测试卡与ELISA法检测HBV的对比分析

目的探讨胶体金测试卡与酶联免疫吸附试验（ELISA）法检测乙型肝炎病毒（HBV）的效果,为临床选择一种经济、快速、准确、敏感检测HBV的新方法。方法分别用胶体金测试卡与ELISA法检测616份血清标本,并对测定结果分别进行比较分析。结果用胶体金测试卡与ELISA法检测乙肝HBsAg、抗-HBs、HBeAg、抗-HBe、抗-HBc,除HbsAg差异无统计学意义外（P〉0.05）,其余差异均有统计学意义（均P〈0.05）。2种方法检测结果HBV5项指标的符合率分别为99.4%、95.3%、98.7%、98.1%、99.0%。以ELISA法为标准验证胶体金测试卡,则胶体金测试卡的灵敏度分别为97.6%、92.4%、88.1%、87.8%、100.0%,特异性分别为100.0%、98.1%、100.0%、100.0%、98.7%。结论胶体金测试卡具有简便、快速、可单份操作的优点,适用于急诊检验和基层检验的需要。但此法与ELISA法相比,其敏感性低,不能单独用于HBV5项定性检测,因此还必须与ELISA法并用。     

Autonomous UAVs for Structural Health Monitoring Using Deep Learning and an Ultrasonic Beacon System with Geo‐Tagging

Visual inspection has traditionally been used for structural health monitoring. However, assessments conducted by trained inspectors or using contact sensors on structures for monitoring are costly and inefficient because of the number of inspectors and sensors required. To date, data acquisition using unmanned aerial vehicles (UAVs) equipped with cameras has become popular, but UAVs require skilled pilots or a global positioning system (GPS) for autonomous flight. Unfortunately, GPS cannot be used by a UAV for autonomous flight near some parts of certain structures (e.g., beneath a bridge), but these are the critical locations that should be inspected to monitor and maintain structural health. To address this difficulty, this article proposes an autonomous UAV method using ultrasonic beacons to replace the role of GPS, a deep convolutional neural network (CNN) for damage detection, and a geo‐tagging method for the localization of damage. Concrete cracks, as an example of structural damage, were successfully detected with 97.7% specificity and 91.9% sensitivity, by processing video data collected from an autonomous UAV.     

呼吸道标本病原学检测对曲霉菌诊断的价值

目的：探讨呼吸道标本直接涂片和培养对曲霉菌诊断的价值。方法将85例呼吸道标本（痰液、肺泡灌洗液）培养曲霉菌阳性患者按曲霉菌感染及曲霉菌定植诊断标准分为曲霉菌感染组（30例）和曲霉菌定植组（55例）。均采用卡方检验和二分类Logistic回归分析细菌培养联合标本直接革兰染色对区别曲霉菌感染和定植的诊断意义。结果85例呼吸道合格标本曲霉菌培养和直接涂片阳性中,30例与临床诊断相符,符合率为35．2％。曲霉菌感染组镜下和培养检出正常菌群均显著低于定植组、曲霉菌检出次数显著高于定植组（均P＜0．05）。2组镜下WBC浸润量、菌丝检出比较差异均无统计学意义（P＝0．408,P＝0．831）。经多因素Logistic回归分析示：曲霉菌检出次数、无正常菌群定植（包括镜下和培养均无正常菌群）对曲霉菌病诊断有统计学意义（P＜0．05）;激素使用、糖尿病、呼吸衰竭及血液系统疾病是其疾病发生的主要影响因素。在曲霉菌培养阳性基础上,镜下未见正常菌群对诊断灵敏度为83．3％,阴性预测值均达83．9％,培养未见正常菌群诊断灵敏度为86．7％、阴性预测值为89．4％,但特异度和阳性预测值均低于60．0％;两者联合诊断曲霉菌感染的特异度可提高到66．4％,阳性预测值可为53．9％。三者联合检测（镜下和培养未见正常菌群与检出次数）能提高曲霉菌诊断的特异度（92．0％）和阳性预测值（71．0％）。结论在标本合格条件下曲霉菌检出阳性时,呼吸道标本直接涂片及培养出正常菌群对区分曲霉菌定植与感染具有诊断价值。     

Evaluation of the lactose Tergitol-7, m-Endo LES, Colilert 18, Readycult Coliforms 100, Water-Check-100, 3M Petrifilm EC and DryCult Coliform test methods for detection of total coliforms and Escherichia coli in water samples

In this study we compared the reference membrane filtration (MF) lactose Tergitol-7 (LTTC) method ISO 9308-1:2000 with the MF m-Endo LES method SFS 3016:2001, the defined substrate chromogenic/fluorogenic Colilert 18, Readycult Coliforms and Water Check methods, and ready-made culture media, 3M Petrifilm EC and DryCult Coli methods for the detection of coliforms and Escherichia coli in various water samples. When the results of E. coli detection were compared between test methods, the highest agreement (both tests negative or positive) with the LTTC method was calculated for the m-Endo LES method (83.6%), followed by Colilert 18 (82.7%), Water-Check (81.8%) and Readycult (78.4%), whereas Petrifilm EC (70.6%) and DryCult Coli (68.9%) showed the weakest agreement. The m-Endo LES method was the only method showing no statistical difference in E. coli counts compared with the LTTC method, whereas the Colilert 18 and Readycult methods gave significantly higher counts for E. coli than the LTTC method. In general, those tests based on the analysis of a 1-ml sample (Petrifilm EC and DryCult Coli) showed weak sensitivity (39.5-52.5%) but high specificity (90.9-78.8%).     

Bacteroidales terminal restriction fragment length polymorphism (TRFLP) for fecal source differentiation in comparison to and in combination with universal bacteria TRFLP

Terminal restriction fragment length polymorphism (TRFLP) is an attractive community analysis method for microbial source tracking (MST) because it is accessible, relatively inexpensive, and can discern multiple fecal sources simultaneously. A new Bacteroidales TRFLP (Bac-TRFLP) method was developed and its source identification performance was evaluated by itself, in comparison to, and in combination with an existing universal bacterial TRFLP method in two laboratories. Sixty-four blind samples from 12 fecal sources (sewage, septage, human, dog, horse, cow, deer, pig, chicken, goose, pigeon, and gull) were used for evaluation. Bac- and Univ-TRFLP exhibited similarly high overall correct identification (>88% and >89%, respectively), excellent specificity regardless of fecal sources, variable sensitivity depending on the source, and stable performance across two laboratories. Compared to Univ-TRFLP, Bac-TRFLP had better sensitivity and specificity with horse, cow, and pig fecal sources but was not suited for certain avian sources such as goose, gull, and pigeon. Combining the general and more targeted TRFLP methods (Univ&Bac-TRFLP) achieved higher overall correct identification (>92%), higher sensitivity and specificity metrics, and higher reproducibility between laboratories. Our results suggest that the Bac-TRFLP and Univ&Bac-TRFLP methods are promising additions to the MST toolbox and warrant further evaluation and utilization in field MST applications.     

Automatic detection method of cracks from concrete surface imagery using two‐step light gradient boosting machine

Automated crack detection based on image processing is widely used when inspecting concrete structures. The existing methods for crack detection are not yet accurate enough due to the difficulty and complexity of the problem; thus, more accurate and practical methods should be developed. This paper proposes an automated crack detection method based on image processing using the light gradient boosting machine (LightGBM), one of the supervised machine learning methods. In supervised machine learning, appropriate features should be identified to obtain accurate results. In crack detection, the pixel values of the target pixels and geometric features of the cracks that occur when they are connected linearly should be considered. This paper proposes a methodology for generating features based on pixel values and geometric shapes in two stages. The accuracy of the proposed methodology is investigated using photos of concrete structures with adverse conditions, such as shadows and dirt. The proposed methodology achieves an accuracy of 99.7%, sensitivity of 75.71%, specificity of 99.9%, precision of 68.2%, and an F‐measure of 0.6952. The experimental results demonstrate that the proposed method can detect cracks with higher performance than the pix2pix‐based approach. Furthermore, the training time is 7.7 times shorter than that of the XGBoost and 2.3 times shorter than that of the pix2pix. The experimental results demonstrate that the proposed method can detect cracks with high accuracy.