关于小麦中呕吐毒素检测质量控制的评价

在同一实验室条件下,通过16名检测人员对小麦中呕吐毒素前处理过程进行人员比对,评价了本实验室在该检测项目的内部质量控制。由长期从事该项目检测的检验人员绘制标准曲线,并通过加标回收率实验验证标准曲线的可靠性。建立免疫亲和层析净化-高效液相色谱法测定小麦中呕吐毒素的含量,在100～5 000 ng/ml的范围内线性关系良好,相关系数为1.000 0,检出限(LOD)为25μg/kg,定量限(LOQ)为83μg/kg;在500、1 000、2 000μg/kg加标水平下,3个浓度水平加标回收率为90.4%～100.1%,3次重复测定的相对标准偏差(RSD)≤2.6%;从16名人员检验数据结果可知,双试验相对标准偏差(RSD)均≤9.4%,Z比分数评价全部均∣Z∣≤1,表明"优秀"。人员比对实验能较好的判定检验人员的检测操作能力与水平,也能加强实验室对检测结果的质量控制,所以应定期进行人员比对实验,为实验室质量管理及质量控制提供重要数据保障。     

食品菌落总数检测人员检测过程与结果比对分析

目的对实验室食品菌落总数检测人员检测过程与结果进行比对,加强实验室质量控制。方法根据内部质量控制的技术要求,按照GB 4789.2-2016《食品微生物学检验菌落总数测定》进行检测,对4名实验人员测定结果进行分析,评价其质量控制。结果 4名检验人员的测定结果均允许范围内,但A人员的测定结果普遍偏低,log值接近于满意范围临界值。结论检验人员严栺觃范地按照有兲操作觃程及标准觃定进行实验,定期进行人员比对,为实验室质量控制及管理提供可靠的数据及保障。     

一种测定食品中二氧化硫含量的质量控制方法

本研究建立一种测定食品中二氧化硫含量的质量控制方法。方法:用碘量法同时标定二氧化硫标准物质和进行加标回收率试验。结果:二氧化硫标准溶液溶度为356.0μg/mL在320～400μg/mL范围内,加标回收率在87.6%～96.3%范围内。结论:该方法快速、方便,适合用于食品中二氧化硫含量检测时的质量控制。     

微滴式数字PCR技术定量检测发酵乳中金黄色葡萄球菌

基于微滴式数字聚合酶链式反应(droplet digital polymerase chain reaction,dd PCR)技术,建立发酵乳中金黄色葡萄球菌定量检测方法。以金黄色葡萄球菌nuc基因为目的片段设计特异性的引物和探针,优化反应体系,通过活菌提取和ddPCR方法对靶标基因的检测特异性和灵敏度进行实验,并对定量结果进行分析。本研究建立了发酵乳中ddPCR技术定量检测金黄色葡萄球菌的方法,检测特异性良好,检测灵敏度为3.3×10~1 CFU/g,定量的偏差率为+10.18%,证明了ddPCR用于绝对定量检测的可行性,为其他食品污染菌和致病菌标准化控制体系提供有益的示范作用。     

超声波、弱酸性电位水在樱桃番茄防腐保鲜中的应用

以樱桃番茄为原料,研究超声波、弱酸性电位水(SAEW)处理对其微生物和货架期的影响。研究发现:超声波、SAEW处理10 min可分别使樱桃番茄表面初始菌落总数降低0.96 log CFU/g和1.45 log CFU/g,霉菌和酵母总数降低了0.68 log CFU/g和1.00 log CFU/g,且在贮藏过程中,处理后的样品表面微生物维持在较低的水平。两种处理手段均能降低樱桃番茄贮藏期的腐烂率和呼吸速率,抑制可溶性固形物含量的下降,而二者对硬度、可滴定酸和维生素C含量无显著影响。本研究结果表明:超声波和电位水在樱桃番茄杀菌防腐,保障其微生物安全性方面具有一定的潜力。     

PCR-核酸试纸条法检测食品中假结核耶尔森菌

目的 建立一种基于PCR-核酸试纸条技术快速检测食品中假结核耶尔森菌的方法。方法 将10株假结核耶尔森菌株和9株其他耶尔森氏菌及18株来源菌株作为实验菌株进行特异性实验; 通过纯菌液计数、干扰菌实验检测进行灵敏度验证。结果 DNA检测可达到10?3 μg/mL, 25 g样品加菌实验灵敏度可达 100 CFU/25 g, 添加10倍干扰菌不会降低检测灵敏度。利用建立方法对市场购买的食品进行筛查并与国标方法进行比较, 建立方法的灵敏度优于国标方法。结论 该方法检测结果准确, 灵敏度高, 适用于检测食品中假结核耶尔森菌。     

食品中致病微生物检测结果的不确定度评定

了解本地区抽查的速冻面米制品的微生物指标状况,并探讨金黄色葡萄球菌的定量检测的数值结果的不确定度评定方法。按照国标法GB 4789.1-2010《食品安全国家标准食品微生物学检验总则》、GB 4789.4-2010《食品安全国家标准食品微生物学检验金黄色葡萄球菌检验》、GB 4789.10-2010《食品安全国家标准食品微生物学检验沙门氏菌检验》和GB 19295-2011《食品安全国家标准速冻面米制品》进行抽样和检测,依据JJF1059.1-2012《测定不确定度评定与表示》,泊松分布及相关统计学方法对检测结果数字不确定度进行评定。抽检的142批次速冻面米制品,两种制品抽检合格率都为100%,得出的两份样品检测结果平均数分别为290 CFU/g和20 CFU/g,相对应该的标准不确定度分别为30 CFU/g和4 CFU/g。本文采用的评定方法,可以对面米制品金黄菌定量检测结果平均数不确定度作出估计,用于速冻面米制品的安全评估。     

食品中金黄色葡萄球菌定量检验能力验证的质量控制

能力验证是利用实验室间比对评价实验室校准或检测能力的重要手段,合理的质量控制是确保能力验证检测结果准确性的关键.本文分别从试剂耗材、操作过程、仪器设备、人员及环境等5个方面对食品中金黄色葡萄球菌定量检测能力验证的影响因素和关键环节进行分析,以期为食品微生物实验室能力验证的质量控制提供参考.     

纺织品实验室化学检测领域内部质量控制的探讨

叙述纺织品实验室化学检测领域的内部质量控制的主要内容,介绍质量控制计划、空白试验、重复检测、比对、加标回收、实验室控制样品和质量控制图等化学检测常用的质量控制方法,结合纺织品化学检测项目,提出内部质量控制的应用指南。     

辣白菜储藏期间病原菌及乳酸菌变化规律的研究

辣白菜在生产过程中易受到有害微生物的污染,并且该产品是一种不经过热加工处理而直接食用的食品,因此残留有害微生物的辣白菜易引发人体痢疾等疾病.本实验将辣白菜在储藏的第1、5、8、12、15、19、22、 26、29、33d分别进行细菌总数、大肠菌群、乳酸菌数、pH值及所含病原菌种类的实验室检测,实验结果表明随着乳酸菌数量的增加,辣白菜中有害微生物数量逐渐降低,到储藏19d时,乳酸菌数量达到1×10 5CFU/g,细菌总数降至1×104CFU/g;到储藏25d时,乳酸菌数量达到1×10 7CFU/g,细菌总数远远小于1×104CFU/g;同时不耐酸性的有害病原菌已经不存在.     

Quantification of Plesiomonas shigelloides in Pure Culture and Clams by Competitive PCR

A quantitative assay for Plesiomonas shigelloides in pure culture and clams based on the competitive polymerase chain reaction was developed. This is the first report for quantitative detection of P. shigelloides by competitive PCR. Forward (PS-F), reverse (PS23RV3), and hybrid primers were designed, and the specificity of the forward and reverse primer for P. shigelloides was proven. An internal standard DNA sequence was synthesized by PCR, with the hybrid primer as the forward primers and PS23RV3 as the reverse primer. A single concentration (0.588 pg/PCR) of internal standard (IS) was used for competitive PCR. The lowest level of detection of P. shigelloides was 80 CFU per PCR in pure culture, 240 CFU/g of clam tissue without enrichment, and 40 CFU/g of clam tissue after 7 hrs. nonselective enrichment at 37°C. There was a linear relationship between the log of the ratio of the relative fluorescent intensities of amplified target DNA bands to the internal standard DNA bands (IS) and the log of the CFU within a certain range in pure cultures and in clam tissue either with or without enrichment. The linear range with cells from a pure culture was the DNA derived from 8.0 × 10¹ to 8.0 × 10⁴ CFU per PCR while with clam tissue, the linear range was the DNA derived from 2.4 × 10² to 2.4 × 10⁵ CFU/g of tissue (1.2 × 10¹ to 1.2 × 10⁴ CFU per PCR) without enrichment, and 4.0 × 10¹ to 1.2 × 10⁴ CFU/g of clam tissue with enrichment, respectively.     

Predictive modelling of the recovery of Listeria monocytogenes on sliced cooked ham after high pressure processing

This study examined bacterial recovery on sliced cooked ham that was inoculated with Listeria monocytogenes, treated by high pressure processing (HPP) and then stored at 10 degrees C for 70 days. The number of L. monocytogenes on the ham inoculated with 5 log(10) CFU/g was initially reduced by HPP at 500 MPa for 10 min to below the detectable level (10 CFU/g). However, the bacterial count gradually increased during storage, and exceeded the initial inoculum level at the end of the 70-day period, having risen by 7-8 log(10) CFU/g. A novel predictive model was therefore developed to estimate the recovery of L. monocytogenes during storage after HPP. Recovery of L. monocytogenes was defined as the detection of >10(2) CFU/g bacteria, in view of the relevant food safety objectives of L. monocytogenes. At each 14-day sampling session, the ham was scored as either 1 or 0 indicating bacterial recovery or no bacterial recovery, respectively. The data were then subjected to a simple linear logistic regression model, which provided a good fit as indicated by the performance statistics. Using this model, we estimated the minimum HPP conditions necessary for the required storage periods. Additionally, as the developed model was based on logistic regression, the probability of the recovery of L. monocytogenes during storage after HPP was estimated. Our model not only calculated the appropriate shelf life and process conditions, but also provided a method for evaluating the risk of the recovery of pathogenic bacteria during storage.     

Rapid and simultaneous quantitation of Escherichia coli 0157:H7, Salmonella, and Shigella in ground beef by multiplex real-time PCR and immunomagnetic separation

The objective of this study was to establish a multiplex real-time PCR for the simultaneous quantitation of Escherichia coli O157:H7, Salmonella, and Shigella. Genomic DNA for the real-time PCR was extracted by the boiling method. Three sets of primers and corresponding TaqMan probes were designed to target these three pathogenic bacteria. Multiplex real-time PCR was performed with TaqMan Universal PCR Master Mix in an ABI Prism 7700 Sequence Detection System. Final standard curves were calculated for each pathogen by plotting the threshold cycle value against the bacterial number (log CFU per milliliter) via linear regression. With optimized conditions, the quantitative detection range of the real-time multiplex PCR for pure cultures was 10(2) to 10(9) CFU/ml for E. coli O157:H7, 10(3) to 10(9) CFU/ml for Salmonella, and 10(1) to 10(8) CFU/ml for Shigella. When the established multiplex real-time PCR system was applied to artificially contaminated ground beef, the detection limit was 10(5) CFU/g for E. coli O157:H7, 10(3) CFU/g for Salmonella, and 10(4) CFU/g for Shigella. Immunomagnetic separation (IMS) was further used to separate E. coli O157:H7 and Salmonella from the beef samples. With the additional use of IMS, the detection limit was 10(3) CFU/g for both pathogens. Results from this study showed that TaqMan real-time PCR, combined with IMS, is potentially an effective method for the rapid and reliable quantitation of E. coli 0157:H7, Salmonella, and Shigella in food.     

Effect of x-ray irradiation on reducing the risk of listeriosis in ready-to-eat vacuum-packaged smoked mullet

Listeria monocytogenes can pose a serious threat in several areas of the nation's food supply including ready-to-eat seafood products. Use of irradiation processing can potentially reduce the risk of listeriosis caused by consumption of ready-to-eat seafood products. This study measured the effect of X-ray irradiation on reducing the population of L. monocytogenes on ready-to-eat, vacuum-packaged smoked mullet. Smoked mullet were inoculated with a five-strain mixture of L. monocytogenes (10(4) CFU/g), vacuum packaged, and irradiated (0, 0.5, 1.0, 1.5, and 2.0 kGy). The packaged fish were then stored at 3 and 10 degrees C for 90 and 17 days, respectively. Radiation doses of 0.5, 1.0, and 1.5 kGy reduced the initial population of L. monocytogenes by 1.1, 1.6, and 2.1 log CFU/g, respectively. The 2.0-kGy dose reduced L. monocytogenes to undetectable levels with no recovery growth at either temperature. Compared to the control, irradiation at 1.5 kGy demonstrated 1.0 and 1.7 log CFU/g less growth at 3 degrees C after 60 days and 10 degrees C after 17 days, respectively. Sensory flavor analysis was conducted to determine if a difference existed between irradiated samples. Panelists indicated that there were no differences among treated and untreated samples. An X-ray dose of 2 kGy effectively eliminated 10(4) CFU/g L. monocytogenes on smoked mullet without changing sensory quality.     

Microbiological quality and safety of ready-to-eat cooked foods from a centralized school kitchen in Argentina

The purpose of this study was to evaluate the microbiological and sensory quality as well as the safety of ready-to-eat (RTE) cooked foods prepared in and distributed from a centralized kitchen to schools in Argentina. A total of 101 cooked food samples delivered as hot RTE cooked foods (group A) and as RTE cooked foods at room temperature (group B) and 140 surface swab environment samples were collected from February to November 1999. Petrifilm plates were used for aerobic (PAC), coliform (PCC), and Escherichia coli (PEC) counts. Standard methods were used to determine Enterobacteriaceae (EntC) and thermotolerant coliform counts (TCC). Samples were also tested for the presence of Salmonella spp., Staphylococcus aureus, Bacillus cereus, and Clostridium perfringens. Food temperatures just before samples were put into containers ranged from 80 to 98 degrees C and from 28 to 32 degrees C for group A and group B, respectively. For group A food samples, PAC ranged from 1.04 to 3.50 log CFU/g, and PCC, PEC, TCC, and EntC were not detected. For group B food samples, PAC ranged from 3.63 to 6.48 log CFU/g, PCC ranged from 1.90 to 5.36 log CFU/g, TCC ranged from 1.30 to 3.95 log CFU/g, and EntC ranged from 3.60 to 5.46 log CFU/g. Of the foodborne pathogens, only B. cereus was isolated (63.4% of samples) in both food groups (<4 log CFU/g). The microbiological and sensory quality and the safety of group A foods were satisfactory. Large numbers of PAC and EntC detected in group B foods show that better control is needed to avoid potential foodborne diseases.     

Evaluation of a 5'-nuclease (TaqMan) assay with the thin agar layer oxyrase method for the detection of Yersinia enterocolitica in ground pork samples

A 5'-nuclease (TaqMan) assay was evaluated for its capability to recover and detect stressed Yersinia enterocolitica. Sensitivity studies of a 5'-nuclease assay for detecting Y. enterocolitica 0:8 in a pure culture system and spiked ground pork samples demonstrated that the assay has reliable sensitivity with a detection limit of 3 to 4 log CFU/ml or CFU/g. The PCR 5'-nuclease (TaqMan) assay was evaluated with the Thin Agar Layer Oxyrase method (TALO, overlaying 14 ml of Trypticase soy agar with a 1:30 dilution of "Oxyrase for Agar" onto a prepoured pathogen-specific, selective medium), and it was compared against the selective medium cefsulodin-irgasan-novobiocin (CIN) for recovering and detecting Y. enterocolitica from inoculated nonfrozen and frozen (-15 degrees C, 2 days) ground pork samples. The TALO method showed more sensitivity (detection limit, 2 log CFU/ml), and it has greater recovery capability (0.5 to 1 log CFU/ml) than CIN (P < 0.05). The 5'-nuclease assay provided rapid detection processing (5 versus 24 h after an 18-h enrichment). The sensitivity per PCR was calculated to as low as 0 to 1 log CFU per PCR reaction; however, in the method's current developmental stage, target pathogens should be enriched to 3 to 4 log CFU/ml or CFU/g to show consistent results. In a survey of 100 ground pork samples using TALO, CIN, and PCR methods, no Y. enterocolitica was recovered. A combined cultivation and an automated PCR TaqMan could be used as a presumptive screening test for detecting Y. enterocolitica in food samples.     

Quantification of Plesiomonas shigelloides in Pure Culture and Clams by Competitive PCR

A quantitative assay for Plesiomonas shigelloides in pure culture and clams based on the competitive polymerase chain reaction was developed. This is the first report for quantitative detection of P. shigelloides by competitive PCR. Forward (PS-F), reverse (PS23RV3), and hybrid primers were designed, and the specificity of the forward and reverse primer for P. shigelloides was proven. An internal standard DNA sequence was synthesized by PCR, with the hybrid primer as the forward primers and PS23RV3 as the reverse primer. A single concentration (0.588 pg/PCR) of internal standard (IS) was used for competitive PCR. The lowest level of detection of P. shigelloides was 80 CFU per PCR in pure culture, 240 CFU/g of clam tissue without enrichment, and 40 CFU/g of clam tissue after 7 hrs. nonselective enrichment at 37°C. There was a linear relationship between the log of the ratio of the relative fluorescent intensities of amplified target DNA bands to the internal standard DNA bands (IS) and the log of the CFU within a certain range in pure cultures and in clam tissue either with or without enrichment. The linear range with cells from a pure culture was the DNA derived from 8.0 × 10¹ to 8.0 × 10⁴ CFU per PCR while with clam tissue, the linear range was the DNA derived from 2.4 × 10² to 2.4 × 10⁵ CFU/g of tissue (1.2 × 10¹ to 1.2 × 10⁴ CFU per PCR) without enrichment, and 4.0 × 10¹ to 1.2 × 10⁴ CFU/g of clam tissue with enrichment, respectively.     

Rapid enumeration of Listeria monocytogenes in artificially contaminated cabbage using real-time polymerase chain reaction

A quantitative real-time polymerase chain reaction (PCR) detection method specific for Listeria monocytogenes was developed, and studies involving pure culture showed that the response of the assay was linear over 7 log10 (log) cycles. The method was then applied to the detection of L. monocytogenes artificially inoculated onto cabbage, a vegetable chosen because it is a major component of coleslaw, which has been associated with an outbreak of listeriosis. After being allowed to attach to the food, cells were washed from the cabbage leaf surface and recovered by centrifugation. The DNA was purified by an organic solvent extraction technique and analyzed by real-time PCR. In this matrix, the method again produced a linear response over 7 log cycles from 1.4 x 10(2) to 1.4 x 10(9) CFU of L. monocytogenes in 25 g of cabbage, and analysis of the reproducibility of the system showed that log differences in L. monocytogenes numbers added to cabbage could be reliably distinguished. The system allowed quantitative results to be obtained within 8 h and was relatively inexpensive, showing good potential for routine analytical use.     

mini-MPN-STE-qPCR法快速定量检测食品中沙门氏菌

目的 建立一种快速、简易、可定量检测食源性沙门氏菌定量PCR(quantitative PCR, qPCR)方法。方法 依据沙门菌属invA基因序列设计染料法qPCR引物, 建立基于嵌合荧光法的沙门氏菌qPCR检测方法, 并通过短时间增菌(short time enrichment, STE)的5管微型多管发酵计数法（mini-most probable number, mini-MPN）进行定量检测, 构建mini-MPN-STE-qPCR法。使用人工污染的鸡肉混合液进行定量检测, 检测结果与传统MPN计数法和平板计数法进行比较分析。结果 建立的 qPCR法特异性良好, 方法灵敏度为50 CFU/mL, 结合48孔板min-MPN和4h 短时间增菌建立的mini-MPN-STE-qPCR法可将整个检测流程缩短至7 h。人工添加沙门氏菌的鸡肉混合液检测结果表明方法检出限为-0.347 log MPN/mL, Bland-Altman分析结果显示, 此法与传统MPN计数法相关系数R2=0.994, 与平板计数法相关系数R2=0.992。结论 该方法准确、简单易行、成本低、灵敏度高, 可用于食品中沙门菌的快速定量检测。