食品中邻苯二甲酸酯类塑化剂检测方法研究进展

塑化剂作为一种加工助剂,可增加聚合物的可塑性,目前已被广泛应用于化工、医药、日用品和食品包装等各领域。邻苯二甲酸酯是最常用的一种塑化剂,其可在生产、流通等过程中迁移到食品内部,对人体造成不可逆伤害。总结了食品中邻苯二甲酸酯类塑化剂检测的前处理方法及传统检测方法的原理、优缺点及适用性,并按照输出信号种类不同分类,综述了几种邻苯二甲酸酯快速检测方法的原理、特点和应用,总结和比较了几种快速检测方法的优缺点,并对其发展方向进行展望,以期为塑化剂检测方法的研究和开发新型快速的检测方法提供参考思路。     

Detection of mutations in human DNA

Efficient methods for the detection of mutations are of fundamental importance in research and in diagnostics. By detection of a DNA sequence alteration that cosegregates with a clinical phenotype in an affected family, the gene at fault may be identified and assigned a function. Mutation detection methods are also a rate-limiting factor for the clinical application of DNA diagnostics. Currently a large number of techniques are in use to scan for new mutations and to distinguish among previously established sequence variants. Here, some of the problems connected with mutation detection are discussed together with principles on which current and future mutation detection assays can be based.     

单核细胞增生李斯特菌检测技术研究进展

单核细胞增生李斯特菌(Listeria monocytogenes,LM)是常见的食源性致病菌之一。目前,在众多单增李斯特菌的检测方法中应用较广的是免疫学检测法、分子学检测法。免疫学检测时间短,操作简单,但该方法依赖高特异性的抗体,会出现假阳性,还需要进一步鉴定检测结果。分子学检测法克服了免疫学检测法不能在种的水平鉴定单增李斯特菌的缺点,省时省力,灵敏度高,但是分子学检测法需要丰富的操作经验,并且不适于现场大批量检测。新兴的代谢学检测法、光谱学检测法、生物传感器等也都有各自的优缺点。本文综合近年最新文献,就单增李斯特菌检测的最新方法、检测进展及未来发展趋势予以分析综述,以期为该菌的检测提供参考。     

食品中单核增生李斯特氏菌检测研究进展

单核增生李斯特氏菌和李斯特菌病的危害近年来引起世界各国食品和卫生部门的广泛关注.关于如何找到一种快速、敏感、准确、合理的检验方法,是当今各国食品卫生部门亟待解决的重要研究课题.对该菌的传统分离方法、免疫学检测方法、核酸检测等方法的最新进展进行了综述,为进行该菌的准确、快速检测奠定了基础.     

An overview of foodborne pathogen detection: In the perspective of biosensors

Food safety is a global health goal and the foodborne diseases take a major crisis on health. Therefore, detection of microbial pathogens in food is the solution to the prevention and recognition of problems related to health and safety. For this reason, a comprehensive literature survey has been carried out aiming to give an overview in the field of foodborne pathogen detection. Conventional and standard bacterial detection methods such as culture and colony counting methods, immunology-based methods and polymerase chain reaction based methods, may take up to several hours or even a few days to yield an answer. Obviously this is inadequate, and recently many researchers are focusing towards the progress of rapid methods. Although new technologies like biosensors show potential approaches, further research and development is essential before biosensors become a real and reliable choice. New bio-molecular techniques for food pathogen detection are being developed to improve the biosensor characteristics such as sensitivity and selectivity, also which is rapid, reliable, effective and suitable for in situ analysis. This paper not only offers an overview in the area of microbial pathogen detection but it also describes the conventional methods, analytical techniques and recent developments in food pathogen detection, identification and quantification, with an emphasis on biosensors.     

炭疽杆菌检测方法的研究现状与展望

炭疽是严重威胁人类健康的烈性传染病,其病原体为炭疽芽孢杆菌。炭疽芽孢杆菌在我国公布的《人间传染的病原微生物名录》中被列为第二类病原微生物(高致病性病原微生物),其芽孢可作为生物战剂和生物恐怖的原材料,因此,发展灵敏、高效的炭疽杆菌检测方法十分重要和紧迫。按检测的靶标分类,针对炭疽杆菌的检测方法主要有四大类:针对炭疽杆菌芽孢的检测方法,针对细菌繁殖体的检测方法,针对炭疽杆菌基因的检测方法和针对炭疽毒素蛋白的检测方法。其中,针对炭疽杆菌芽孢和细菌繁殖体的检测已经有比较成熟的方法,但其在特异性以及临床的实用性方面难以令人满意;针对炭疽杆菌基因的检测技术在特异性和灵敏度上有较大的提高,但在临床诊断等方面还有欠缺;而针对炭疽毒素蛋白的检测技术的发展,使得直接对炭疽杆菌的主要致病因子的检测成为可能,这对于临床诊断以及流行病学研究具有重要意义。本文对当前炭疽杆菌检测方法的最新进展做了简要的归纳,关注了不同检测方法的适用范围和检测能力,并展望了相关领域的发展趋势,希望能为从事炭疽杆菌检测方法研究的同行提供参考和帮助。     

Potentials and limitations of molecular diagnostic methods in food safety

Molecular methods allow the detection of pathogen nucleic acids (DNA and RNA) and, therefore, the detection of contamination in food is carried out with high selectivity and rapidity. In the last 2 decades molecular methods have accompanied traditional diagnostic methods in routine pathogen detection, and might replace them in the upcoming future. In this review the implementation in diagnostics of four of the most used molecular techniques (PCR, NASBA, microarray, LDR) are described and compared, highlighting advantages and limitations of each of them. Drawbacks of molecular methods with regard to traditional ones and the difficulties encountered in pathogen detection from food or clinical specimen are also discussed. Moreover, criteria for the choice of the target sequence for a secure detection and classification of pathogens and possible developments in molecular diagnostics are also proposed.     

High-performance liquid chromatographic separation and detection methods for anabolic compounds

The role of high-performance liquid chromatography (HPLC) in methods of analysis for anabolic compounds in biological samples is reviewed. Special attention is given to both the separation and detection of anabolic compounds. A distinction is made between on-line detection systems, such as ultraviolet detection and diode-array detection, and off-line detection methods with special emphasis on immunochemical detection methods using non-isotopic labels. A number of applications are given to elucidate the possibilities of HPLC in the analysis of anabolic compounds.     

基于纳米金的比色分析法在核酸检测中的应用

随着纳米技术的发展,运用纳米粒子检测核酸成为研究的热点.在众多检测方法中,基于纳米金的比色分析法操作较为简便,只需普通光学仪器甚至肉眼即可观察结果,从而表现出广阔的市场及临床应用前景.基于纳米金的比色分析法有多种,不同检测原理的方法在灵敏度和实用性上存在差异.根据纳米金是否经寡核苷酸探针修饰可将其分为基于功能化纳米金的比色分析法和基于未功能化纳米金的比色分析法,前者又分为利用纳米金颜色变化的聚集反应体系以及利用纳米金特殊氧化-还原能力的银染增强体系.     

A low-cost, portable generic biotoxicity assay for environmental monitoring applications

Fish chromatophores have been shown to be promising biosensors for the detection of hostile agents in the environment. However, state-of-art methods for such applications are still based on extensive use of data/signal processing, in conjunction with need for a skilled human observer to carry out the detection. As a result, conventional methods are complex, costly and cumbersome rendering them useless for field applications requiring low-cost portable solutions capable of fast detection. A new technique is proposed based on the popular scheme of observing the aggregation response in chromatophores for detection of toxicity, and a solution using optical detection and electronic processing is outlined. This scheme has the advantage of being low in cost while providing simple, fast and reliable detection.     

Sentinel lymph node detection by combining nonradioactive techniques with contrast agents: State of the art and prospects

The status of sentinel lymph nodes (SLNs) has a substantial prognostic value because these nodes are the first place where cancer cells accumulate along their spreading route. Routine SLN biopsy (“gold standard”) involves peritumoral injections of radiopharmaceuticals, such as technetium-99m, which has obvious disadvantages. This review examines the methods used as “gold standard” analogs to diagnose SLNs. Nonradioactive preoperative and intraoperative methods of SLN detection are analyzed. Promising photonic tools for SLNs detection are reviewed, including NIR-I/NIR-II fluorescence imaging, photoswitching dyes for SLN detection, in vivo photoacoustic detection, imaging and biopsy of SLNs. Also are discussed methods of SLN detection by magnetic resonance imaging, ultrasonic imaging systems including as combined with photoacoustic imaging, and methods based on the magnetometer-aided detection of superparamagnetic nanoparticles. The advantages and disadvantages of nonradioactive SLN-detection methods are shown. The review concludes with prospects for the use of conservative diagnostic methods in combination with photonic tools.     

Identification and quantification of polyphenol phytoestrogens in foods and human biological fluids

We review the methods used to measure phytoestrogens (genistein, daidzein, lignans and their derivatives) in foods and biological fluids, and discuss advantages and disadvantages of each. The range of detection limits reported varies widely between individual laboratories, but generally the best reported sensitivity is as follows: immunoassay>HPLC-mass spectrometry=HPLC-multichannel electrochemical detection (coularray)>GC-single ion monitoring-mass spectrometry>HPLC-UV diode array>HPLC-single channel electrochemical detection. The best sensitivity reported so far is 0.002 pmol per assay for daidzein by radioimmunoassay. HPLC with UV diode array detection is the most commonly employed, but is the least sensitive and specific. GC and HPLC coupled with mass spectrometry or electrochemical detection are the most accurate and reproducible methods for a wide variety of analytes. Generally most methods, with the exception of immunoassay, have not been correlated with other methods. Recoveries from extraction methods, limits of detection, nature of compounds analysed and the internal standards used are summarised for more than 90 reports in the literature. From this data, it is clear that an inter-laboratory validation and correlation between a wide range of methods for phytoestrogen analysis is required. One underdeveloped area that requires particular attention is the analysis of plant lignans.     

Detection and enumeration of coliforms in drinking water: current methods and emerging approaches.

The coliform group has been used extensively as an indicator of water quality and has historically led to the public health protection concept. The aim of this review is to examine methods currently in use or which can be proposed for the monitoring of coliforms in drinking water. Actually, the need for more rapid, sensitive and specific tests is essential in the water industry. Routine and widely accepted techniques are discussed, as are methods which have emerged from recent research developments.Approved traditional methods for coliform detection include the multiple-tube fermentation (MTF) technique and the membrane filter (MF) technique using different specific media and incubation conditions. These methods have limitations, however, such as duration of incubation, antagonistic organism interference, lack of specificity and poor detection of slow-growing or viable but non-culturable (VBNC) microorganisms. Nowadays, the simple and inexpensive membrane filter technique is the most widely used method for routine enumeration of coliforms in drinking water.The detection of coliforms based on specific enzymatic activity has improved the sensitivity of these methods. The enzymes beta-D galactosidase and beta-D glucuronidase are widely used for the detection and enumeration of total coliforms and Escherichia coli, respectively. Many chromogenic and fluorogenic substrates exist for the specific detection of these enzymatic activities, and various commercial tests based on these substrates are available. Numerous comparisons have shown these tests may be a suitable alternative to the classical techniques. They are, however, more expensive, and the incubation time, even though reduced, remains too long for same-day results. More sophisticated analytical tools such as solid phase cytometry can be employed to decrease the time needed for the detection of bacterial enzymatic activities, with a low detection threshold.Detection of coliforms by molecular methods is also proposed, as these methods allow for very specific and rapid detection without the need for a cultivation step. Three molecular-based methods are evaluated here: the immunological, polymerase chain reaction (PCR) and in-situ hybridization (ISH) techniques. In the immunological approach, various antibodies against coliform bacteria have been produced, but the application of this technique often showed low antibody specificity. PCR can be used to detect coliform bacteria by means of signal amplification: DNA sequence coding for the lacZ gene (beta-galactosidase gene) and the uidA gene (beta-D glucuronidase gene) has been used to detect total coliforms and E. coli, respectively. However, quantification with PCR is still lacking in precision and necessitates extensive laboratory work. The FISH technique involves the use of oligonucleotide probes to detect complementary sequences inside specific cells. Oligonucleotide probes designed specifically for regions of the 16S RNA molecules of Enterobacteriaceae can be used for microbiological quality control of drinking water samples. FISH should be an interesting viable alternative to the conventional culture methods for the detection of coliforms in drinking water, as it provides quantitative data in a fairly short period of time (6 to 8 h), but still requires research effort.This review shows that even though many innovative bacterial detection methods have been developed, few have the potential for becoming a standardized method for the detection of coliforms in drinking water samples.     

土地覆盖变化检测方法比较——以内蒙古草原区为例

随着对地观测技术的不断发展,遥感影像分辨率逐渐提高,促进了基于遥感影像的变化检测从传统像元级的检测向面向对象的检测转变。为了探究面向对象的变化检测方法在土地覆盖变化检测中的有效性和适用性,对面向对象的变化检测方法与常规的变化检测方法进行对比评价。以内蒙古鄂尔多斯和包头地区为试验区,选取2002年及2011年的Landsat TM/ETM+影像为数据源,比较了图像代数运算、图像变换、图像空间结构特征和面向对象的多种变化检测方法,对研究区两期土地覆盖进行了变化检测研究。结果表明:面向对象的变化检测方法在总体精度、kappa系数上都有明显的优越性,总体精度均在87.42%以上,尤其以面向对象的变化矢量分析方法精度最高,达91.56%。此外,主成分差异法也有较好的检测效果,总体精度为87.83%。对总体精度较高的3种方法在不同土地覆盖变化类型中检测效果的研究表明:对于研究区几种主要土地覆盖变化类型,面向对象的变化矢量分析法均有较理想的检测效果,平均精度为85%左右,且始终优于面向对象的光谱向量相似法,以居民地及旱地相关的变化类型最为明显;主成分差异法对不同土地覆盖变化类型检测效果差异很大,对其中4种变化类型的精度甚至达到了93%以上,但对于检测草地与裸地间转化精度很低,甚至只有8.69%;在与工矿用地有关的土地覆盖变化类型中,面向对象的变化矢量分析法的精度明显高于主成分差异法,而在与居民地有关的变化类型中,主成分差异法表现出一定优势。     

Comparison of a cultural method with ListerScreen plus Rapid'L.mono or PCR-ELISA methods for the enumeration of L. monocytogenes in naturally contaminated sewage sludge

Cultural methods used to count Listeria monocytogenes in sewage sludge are laborious and time consuming, and alternative methods are needed to reduce analysis time and improve detection limits. In this study, a survey of L. monocytogenes in sewage sludge is presented with a comparative study between a cultural method and immunomagnetic separation using a ListerScreen test followed by identification of L. monocytogenes with Rapid'L.mono agar or PCR-ELISA. These two alternative methods improved the detection of L. monocytogenes in different types of sludge, irrespective of their physical and chemical characteristics. The ListerScreen method coupled with detection of L. monocytogenes on Rapid'L.mono offers the advantage of being less sophisticated than the molecular method and allows isolation of the organism, which may be useful in epidemiological studies. However, ListerScreen coupled with PCR-ELISA proved best for high-sensitivity detection of L. monocytogenes in sewage samples.     

Spatial Cluster Detection for Repeatedly Measured Outcomes while Accounting for Residential History

Spatial cluster detection has become an important methodology in quantifying the effect of hazardous exposures. Previous methods have focused on cross‐sectional outcomes that are binary or continuous. There are virtually no spatial cluster detection methods proposed for longitudinal outcomes. This paper proposes a new spatial cluster detection method for repeated outcomes using cumulative geographic residuals. A major advantage of this method is its ability to readily incorporate information on study participants relocation, which most cluster detection statistics cannot. Application of these methods will be illustrated by the Home Allergens and Asthma prospective cohort study analyzing the relationship between environmental exposures and repeated measured outcome, occurrence of wheeze in the last 6 months, while taking into account mobile locations.     

Evaluation of non-radioactive labelling and detection of deoxyribonucleic acids: Part one: chemiluminescent methods

The growth of analytical methods for the detection of nucleic acid from various biological samples reflects recent advances in biotechnology development especially in the areas of genetic, infections and cancer diagnosis. The target DNA is detected by hybridization techniques derived from Southern's blotting. However such assays, based on the use of ³²P labelled DNA probes, bring with them the associated problems of handling radioactive materials. In order to overcome these difficulties, a number of chemiluminescent detection methods have recently been developed.These new, alternative probe labelling procedures and chemiluminescent detection methods are easy to use in routine assays performed in research laboratories as well as for medical applications, and can reach the level of sensitivity found in classical radiolabelling techniques.The techniques investigated include peroxydase, biotin 16-dUTP or digoxigenin 11-dUTP probe labelling. The target DNAs are transferred onto nitrocellulose or nylon membranes and further fixed by heat or UV crosslinking. Specific hybridization on the target DNA is finally revealed by the use of chemiluminescent substrates. For all these techniques the detection limit is 10 aM (attomol) of a 561 bp target DNA. However for the probes labelled with peroxydase and with digoxigenin the detection limit drops to 1.0 aM of the target DNA. In the present paper we shall compare several of these DNA labelling and detection procedures and show that the detection threshold can vary by as much as a factor of 20 from method to method. This is the first time that various chemiluminescent methods for label and detection of DNA are compared and evaluated in order to determine the best protocol.     

Balancing Precision and Risk: Should Multiple Detection Methods Be Analyzed Separately in N-Mixture Models?

Using multiple detection methods can increase the number, kind, and distribution of individuals sampled, which may increase accuracy and precision and reduce cost of population abundance estimates. However, when variables influencing abundance are of interest, if individuals detected via different methods are influenced by the landscape differently, separate analysis of multiple detection methods may be more appropriate. We evaluated the effects of combining two detection methods on the identification of variables important to local abundance using detections of grizzly bears with hair traps (systematic) and bear rubs (opportunistic). We used hierarchical abundance models (N-mixture models) with separate model components for each detection method. If both methods sample the same population, the use of either data set alone should (1) lead to the selection of the same variables as important and (2) provide similar estimates of relative local abundance. We hypothesized that the inclusion of 2 detection methods versus either method alone should (3) yield more support for variables identified in single method analyses (i.e. fewer variables and models with greater weight), and (4) improve precision of covariate estimates for variables selected in both separate and combined analyses because sample size is larger. As expected, joint analysis of both methods increased precision as well as certainty in variable and model selection. However, the single-method analyses identified different variables and the resulting predicted abundances had different spatial distributions. We recommend comparing single-method and jointly modeled results to identify the presence of individual heterogeneity between detection methods in N-mixture models, along with consideration of detection probabilities, correlations among variables, and tolerance to risk of failing to identify variables important to a subset of the population. The benefits of increased precision should be weighed against those risks. The analysis framework presented here will be useful for other species exhibiting heterogeneity by detection method.     

Determination of polyhydroxyalkanoates in activated sludge by ion chromatographic and enzymatic methods

Two new detection methods for the determination of poly-beta-hydroxybutyrate (PHB) and -valerate (PHV) are described. Both methods are based on depolymerization of PHB/PHV to 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV). Depolymerization was achieved by either propanolic or hydrolytic digestion. Propanolic digestion transformed commercial PHB/PHV stoichiometrically into 3HB/3HV and yielded apparently complete recoveries of bacterial PHB/PHV from activated sludge. Hydrolytic digestion was suitable only for PHB determination. For quantification of 3HB and 3HV directly from digested sludge, a method based on ion-exchange chromatography and conductivity detection was developed (IC-method). Alternatively, the total of 3HB and 3HV was quantified using a commercial enzymatic test kit and colorimetric detection (enzyme method). Both detection methods are easier to perform than previous methods and are suitable for complex matrices such as activated sludge. The IC-method is recommended for high sample throughputs or if distinction between PHB and PHV is essential. Enzymatic detection is recommended if a few samples per day have to be measured immediately or if an ion chromatograph is unavailable.     

Line Transect Sampling in Small Regions

This article develops an approach to estimating population abundance from line transect surveys that uses a calibration survey to estimate the detection function, which is then employed as a weight function in constructing the abundance estimate. Nonparametric methods of estimating the detection function via local regression and via a kernel density estimator are considered. The proposed methods are evaluated using a set of Western Australian plant data and weed enumeration data.