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。     

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。     

PCR技术在梅毒检测中的应用

梅毒是一种由梅毒螺旋体(TP)感染引起的慢性性传播疾病,其临床表现复杂多样。目前常用实验室诊断方法主要有暗视野显微镜法和血清学方法。近年来,聚合酶链式反应(PCR)技术在梅毒实验室诊断中得以应用,在早期梅毒以及特殊梅毒的诊断中显示出灵敏度高、特异性较强、稳定性好等优点。本文就PCR检测技术在梅毒实验室诊断中的研究进展予以综述。     

透水混凝土孔隙率快速检测方法

透水混凝土因具有多孔结构的特性而具有透水透气的功能特性。但目前没有针对新拌透水混凝土材料的孔隙率现场检测方法。针对这种现状,通过对透水混凝土的结构模型分析和试验设计找出了适用于透水混凝土现场质量检测的一种方法。该方法通过与国内采用重量法分析方法对比分析得出,透水混凝土快速检测方法数据可靠,与已有方法的测量结果拟合度高。并且该方法具有及时检测的优点,对透水混凝土结构物的施工质量控制有实际指导意义。     

水泥强度快速检测方法评述

在水泥生产和工程施工中,快速检测水泥强度存在着很大的必要性。本文将对水泥快速检测强度方法种类的归纳,分析其方法机理、应用范畴等,为生产和施工企业快速检测水泥强度带来一定的借鉴。     

透水混凝土孔隙率快速检测方法

透水混凝土因具有多孔结构的特性而具有透水透气的功能特性。但目前没有针对新拌透水混凝土材料的孔隙率现场检测方法。针对这种现状,通过对透水混凝土的结构模型分析和试验设计找出了适用于透水混凝土现场质量检测的一种方法。该方法通过与国内采用重量法分析方法对比分析得出,透水混凝土快速检测方法数据可靠,与已有方法的测量结果拟合度高。并且该方法具有及时检测的优点,对透水混凝土结构物的施工质量控制有实际指导意义。     

食品中重金属元素检测方法研究进展

食品安全问题是直接关系到人民健康的重大民生问题。简要阐述了食品中重金属对人体的影响与危害,全面分析了重金属元素对食品的污染,从不同种类的食品,包括农作物(粮食、蔬菜、水果、食用菌、茶叶)、水产品、畜禽产品、食用油、食用酒精等方面,综述了近年来食品中重金属检测方法研究的进展。随着检测技术的不断发展进步,快速、灵敏、无损的多元素在线检测的方法必将在食品中重金属元素的测定方面得到广泛的应用。     

食品中重金属检测方法研究探讨

如今重金属污染问题已经日趋严重,在食品、医药、环境等诸多方面都引起了人们广泛的重视,近年来重金属快速检测的方法发展迅速,在重金属快速检测与筛查方面发挥了很大的作用,但还需进一步改进和完善。目前,重金属检测仍然存在以下不足,忽略了对有益重金属的测定,忽略了对重金属元素的不同价态分别进行测定,还有不容忽视样品中其它物质对测定的影响。本论文从应用新型的科技成果到结合生物技术对重金属的检测方法做出探讨。     

沥青红外光谱快速检测方法探讨

沥青在道路建设中是一种常用材料,主要分为基质沥青、改性沥青等。利用沥青针入度、软化点、延度、蜡含量和SBS含量等指标,可以将沥青的性能充分体现出来,这些指标是沥青生产、销售和使用等环节需要重点检测的。因此,为了保证沥青性能满足使用条件,文章将介绍沥青红外光谱快速检测的方法。     

用PCR技术检测水中隐孢子虫

介绍了一种检测水中隐孢子虫的巢式PCR方法：根据Cryptosporidium parvum 18s rRNA基因序列选用了两对引物，特异性扩增了该基因可变区中1056bp和435bp目标片段。加标试验表明，该方法的检测限为10个卵囊。     

水环境中微生物PCR检侧灵敏度的研究

以大肠杆菌纯培养物和清华大学湖水以及自来水中的微生物为研究对象,研究了不同水环境中微生物PCR检刚的检出限,结果表明:大肠杆菌JM 109纯培养物的PCR检出限为2.6x10～3CFU/mL,湖水的PCR检出限为1.2x10～3CFU/mL,自来水中微生物的PCR检出限为2.3x10～3CFU/mL.湖水和自来水中的检出限均低于大肠杆菌纯培养物.     

N-terminal amino acid sequence identity between a major allergen of Ascaris lumbricoides and Ascaris suum,and MHC-restricted IgE responses to it

A protein allergen of the parasitic nematode Ascaris has been purified to homogeneity by immunoaffinity chromatography. It is the most abundant protein species in the parasite''s body fluid and has been named ABA-1. The allergen''s molecular weight (MW) has been previously estimated at 14,000, but this sizing is currently under re-evaluation. The immunological activity of the protein was intact after purification, as attested by immunoprecipitation and passive cutaneous anaphylaxis. The IgE response to ABA-1 was under major histocompatibility complex (MHC) restriction in the rat, in which only RT1u strains were found to respond following infection with the parasite. The tissue-invasive and intestinal stages of both Ascaris lumbricoides (of humans) and Ascaris suum (of pigs) have an antigen of similar MW to ABA-1 in their secretions or among their somatic antigens. These are antigenically indistinguishable; they were found to have similar amino acid compositions, and their N-terminal amino acid sequences were identical to 41 residues. Finally, the apparent MW, amino acid composition and isoelectric point of ABA-1 all argue for close similarity to the previously described Allergen A of the parasite.     

Detection of viable bacteria in environmental water samples using DNase I and PCR method

In this study, we tested the potential application of a previously developed method in detecting Escherichia coli in environmental water samples. To increase the sensitivity of the method, and the recovery of microbial cells, water samples were filtered before being subjected to DNase treatment and polymerase chain reaction amplification. Results showed that DNase I treatment and PCR reaction were not affected by inhibitors as the expected amplicon was successfully amplified in autoclaved environmental waters spiked with E. coli. Then, we applied this method to naturally contaminated environmental water samples. We firstly confirmed the presence of coliforms and E. coli in these water samples by plating in eosin methylene blue agar. Simultaneous PCR amplification targeting Lac Z and uidR gene of total coliforms and E. coli respectively demonstrated that this developed method is potentially applicable for routine microbial assessment of health risks related to viable microorganisms in environmental or drinking waters.     

Precision and accuracy of an assay for detecting Ascaris eggs in various biosolid matrices

This paper presents quality assurance data and quality control data on the recovery of Ascaris suum eggs from various biosolid matrices: acid-treated, alkaline-treated, amended-soil blended, and lagoon stored biosolids. Over a period of years, the same procedure, the "Tulane Method," was performed on different matrices, and in this work, the data collected on the recovery of the eggs from the different matrices is examined and compared. The egg recoveries are discussed in terms of precision (the comparison of the recovery of eggs from a sample processed in duplicate) and in terms of accuracy (the percentage of eggs recovered from a sample to which eggs were added at the beginning of the extraction procedure). This form of quality analysis/control is typically called the "Split/Spike" method. This method of biosolid processing for helminth egg recovery had an overall accuracy of about 60% or greater and a percent variation from the mean density (an indirect method of assessing precision) of only 3-35%.     

Quantification of anaerobic ammonium-oxidizing bacteria in enrichment cultures by real-time PCR

The anaerobic ammonium-oxidizing (ANAMMOX) bacteria were enriched from a rotating disk reactor (RDR) biofilm in semi-batch cultures. Based on fluorescence in situ hybridization (FISH) analysis, this enrichment led to a relative population size of 36% ANAMMOX bacteria. Phylogenetic analysis revealed that all the detected clones were related to the previously reported ANAMMOX bacteria, Candidatus Brocadia anammoxidans (AF375994), with 92% sequence similarity. Furthermore, we successfully developed a real-time polymerase chain reaction (PCR) assay to quantify populations of ANAMMOX bacteria in the enrichment cultures. For this real-time PCR assay, PCR primer sets targeting 16S ribosomal RNA genes of ANAMMOX bacteria were designed and used. The quantification range of this assay was 6 orders of magnitude, from 8.9x10(1) to 8.9x10(6) copies per PCR, corresponding to the detection limit of 3.6x10(3) target copies mL(-1). A significant correlation was found between the increase in copy numbers of 16S rRNA gene of ANAMMOX bacteria and the increase in nitrogen removal rates in the enrichment cultures. Quantifying ANAMMOX bacterial populations in the enrichment culture made it possible to estimate the doubling time of the enriched ANAMMOX bacteria to be 3.6 to 5.4 days. The real-time PCR assay gave comparable population sizes in the enrichment cultures with the FISH results. These results suggest that the real-time PCR assay developed in this study is useful and reliable for quantifying the populations of ANAMMOX bacteria in environmental and engineering samples.     

聚合酶链式反应在微生物检测中的应用

综述了聚合酶链式反应（PCR技术）的原理、方法及其研究进展，介绍了PCR及其相关技术在水体微生物检测中的优势和应用研究现状，分析了该技术在水体微生物检测中存在的问题及解决方法。PCR技术的应用将极大地提高水体微生物检测速度、灵敏度和准确度，对于评价水处理过程中细菌、病毒、病原原生生物等的去除效率也将具有重要意义。     

基于PCR的区域物流综合发展水平影响因素实证研究——以江苏省为例

区域物流极大地促进了区域经济的增长．带动了周边地区的发展。通过对江苏省区域物流的研究利用主成分回归的方法．构建了影响区域物流综合发展水平的回归模型。经回归分析得出,社会经济的发展,产业结构的合理化布局、物资需求流通的加速、物流基础设施的优化．以及信息技术的及时跟进对区域物流的发展起到了正向促进作用。尤其是经济的快速发展、产业结构合理化布局,物资流通的加速、公路设施的完善、邮政服务水平的提升．更是对区域物流的发展起到了至关重要的作用。