金黄色葡萄球菌TraP蛋白单克隆抗体制备

研究通过骨髓瘤细胞SP2／O与经金黄色葡萄球菌（S．aureus）Trap蛋白免疫的小鼠脾细胞融合,并进一步筛选种单克隆化,获得了11株稳定分泌抗TraP蛋白的单克隆抗体（McAb）杂交瘤细胞株,其中8株McAb（2A1、3A6、381、184、2C5、4C7、5C3和2D8）亚类为IgGl,3株McAb（2A7、3A1和1c4）亚类为IgM,轻链均为K链。8株IgGl型McAb的小鼠腹水的抗体效价均达到1：128000,交叉实验结果显示这8株McAb不与S．aureus的ClfA、ClfB、Cna、FnbPA和IsdB蛋白以及链球菌的GapC蛋白反应,具有良好的特异性,Western—blotting结果显示这8株McAb识别的都是TraP的线性表位。     

Antimicrobial activity of enrofloxacin against Staphylococcus intermedius strains isolated from canine pyodermas

This study examined and compared the minimal inhibition concentrations (MICs) of enrofloxacin against 393 Staphylococcus intermedius strains isolated in France from canine pyodermas during three different years, 1995 (174 isolates), 1997 (101 isolates) and 1999 (118 isolates). The MICs of enrofloxacin against these strains ranged from 0.063 to 64 mg L^?1, with MIC₅₀ and MIC₉₀ equal to 0.125 and 0.25 mg L^?1, respectively. Two resistant strains were found, but only among isolates collected in 1999. The data show that resistance to enrofloxacin among S. intermedius strains is still rare in dogs, but the selection in vitro of variants in which the MICs were increased 4–16‐fold after 10 serial passages in subinhibitory concentrations of enrofloxacin suggests that inappropriate use might favour the development of resistant strains in vivo.     

A model to determine sampling strategies and milk inoculum volume for detection of intramammary Staphylococcus aureus infections in dairy cattle by bacteriological culture

A model was developed to evaluate the effects that methods of obtaining milk samples and culture inoculum volumes had on the sensitivity of microbiological culture to detect Staphylococcus aureus intramammary infections (IMI). An assumption was made that milk from mammary quarters infected with S. aureus only contains bacteria intermittently. A modified sine wave function was used to model this intermittent shedding pattern. Specifications for the components of the shedding cycle used in this function were based on quantitative culture results from 54 experimentally infected S. aureus quarters, sampled daily for a period of 30–49 days. The components of the shedding cycle were length in days, peak number of CFU shed per milliliter of milk, and length of time in the cycle when no shedding occurred. These components were used to estimate the model's predicted distribution of S. aureus CFU ml⁻¹ milk when individual quarter milk samples were cultured for S. aureus. The sensitivity of culture for several sampling methods was then calculated. The model predicted that culture of a single quarter milk sample had a sensitivity ranging from 60 to 87% for detection of S. aureus IMI depending on inoculum volume. Quarter milk samples taken on day 1 and repeated either on day 3 or day 4, and cultured separately using 0.1 ml of milk for culture inoculum, were predicted to have sensitivities of 90–95% and 94–99%, respectively. Other milk-sampling strategies examined included culture of a composite milk sample (equal-volume mixture of milk from four separate mammary quarters ) and pooled milk samples in which samples from different milkings (either quarter or composite samples) were mixed together and then cultured. The range of predicted sensitivities of these other sampling strategies was 30–97%. Factors having the greatest impact on the sensitivity of culture, in order of importance were: the type of milk sample, the volume of milk cultured, and the time interval between repeated milk sample collection strategies.     

奶牛乳房炎病例中金黄色葡萄球菌RAPD-PCR基因分型的研究

用随机扩增多态性DNA 技术（RAPD）对来自2个集约化奶牛场不同奶牛乳房炎病例分出的16株金黄色葡萄球菌进行了基因分型。分别用3个寡核苷酸引物扩增，仅OLP11引物可扩增出质量良好、具有RAPD特征的条带图谱。对反应体系的退火温度、模板浓度进行了筛选，模板原倍浓度、33℃的退火温度能使所有检测菌株产生清晰、可分辨、具有较好重复性的带谱。分型结果表明，16株菌可分为2个类群，其中14株位于同一类群，大多数菌株的相关系数差距仅为0.04，这一结果提示这些菌株可能具有相同的来源和相似的遗传背景。     

PCR-based detection of genes encoding virulence determinants in Staphylococcus aureus from bovine subclinical mastitis cases

The present study was carried out to genotypically characterize Staphylococcus aureus (S. aureus) isolated from bovine mastitis cases. A total of 37 strains of S. aureus were isolated during processing of 552 milk samples from 140 cows. The S. aureus strains were characterized phenotypically, and were further characterized genotypically by polymerase chain reaction using oligonucleotide primers that amplified genes encoding coagulase (coa), clumping factor (clfA), thermonuclease (nuc), enterotoxin A (entA), and the gene segments encoding the immunoglobulin G binding region and the X region of protein A gene spa. All of the isolates yielded an amplicon with a size of approximately 1,042 bp of the clfA gene. The amplification of the polymorphic spa gene segment encoding the immunoglobulin G binding region was observed in 34 isolates and X-region binding was detected in 26 isolates. Amplification of the coa gene yielded three different products in 20, 10, and 7 isolates. The amplification of the thermonuclease gene, nuc, was observed in 36 out of 37 isolates. All of the samples were negative for the entA gene. The phenotypic and genotypic findings of the present strategies might provide an understanding of the distribution of the prevalent S. aureus clones among bovine mastitis isolates, and might aid in the development of steps to control S. aureus infections in dairy herds.     

病毒细菌协同凝集试验在蔬菜病毒检测中的应用

在蔬菜病毒病的病毒细菌协同凝集试验（简称ＶＢＡ法）中,首次比较了金黄色葡萄球菌Ｃｏｍｎ１和Ｎｏ．１８００菌株对ＶＢＡ灵敏度的影响,结果表明用Ｎｏ．１８００菌株代替国内、外通用的Ｃｏｍｎ１菌株,ＶＢＡ检测ＴｏＭＶ、ＣＭＶ和ＰＶＸ纯病毒的灵敏度提高了２～１０倍,达３．４～１３．０ｎｇ／ｍＬ,检出病汁液的最大稀释度达１０３～１０５,其灵敏度接近于Ａ蛋白酶联法。在此基础上,又将经抗血清致敏的金黄色葡萄球菌液体免疫剂冷冻干燥,制成一种新型的固体免疫剂,可在４℃下保存;在－１０℃贮藏１年后,用ＶＢＡ法检测ＴｏＭＶ、ＣＭＶ和ＰＶＸ纯病毒的灵敏度分别为８１、２０５和２１０ｎｇ／ｍＬ;即使在１５℃下贮藏１年,灵敏度仍高达２～６μｇ／ｍＬ;在２５℃下贮藏３个月,还有一定的效价。而液体免疫剂不能在１５℃下贮藏;在４℃下贮藏１０个月即失效。因此,固体免疫剂既保持了原液体免疫剂灵敏度高、特异性强、快速简便和经济的特点,又增加了耐贮藏的优点,更加适合在基层单位推广使用。     

Evaluation of prevalence and risk factors for methicillin-resistant Staphylococcus aureus colonization in veterinary personnel attending an international equine veterinary conference

Infection and colonization with methicillin-resistant Staphylococcus aureus (MRSA) are now increasingly being reported in animals and people who work with animals, including horses and horse personnel. The objectives of this study were to evaluate the prevalence of MRSA colonization in veterinary personnel attending an international equine veterinary conference, and to identify risk factors for MRSA colonization in this group. Volunteers were recruited at the conference to provide a single nasal swab and complete a questionnaire regarding contact with the human health care system, contact with horses, and personal hand hygiene. Colonization with MRSA was found in 26/257 (10.1%) of study participants. Multivariable analysis showed an increased risk of MRSA colonization associated with having been diagnosed with or having treated a patient diagnosed with MRSA colonization or infection in the last year (odds ratio [OR] 8.41, 95% confidence interval [CI] 1.51–49.92, P = 0.015; OR 3.27, 95% CI 1.05–10.11, P = 0.039, respectively), whereas hand washing between infectious cases (OR 0.27, 95% CI 0.11–0.72, P = 0.009) and hand washing between farms (OR 0.35, 95% CI 0.14–0.98, P = 0.047) were protective. The prevalence of MRSA colonization among equine veterinary personnel found in this study was high compared to that of other studies of the general population. These data support previous suggestions that equine veterinary personnel are at increased risk of colonization with MRSA. This is the first study to demonstrate a statistically significant association between hand hygiene practices and a measurable clinical outcome in veterinary medicine.     

鸡外周血白细胞吞噬试验的研究

[目的]确定鸡外周血白细胞吞噬葡萄球菌的最佳条件。[方法]使用葡萄球菌10145为吞噬材料,以瑞士染色法检测鸡外周血白细胞的吞噬活性。通过正交试验研究吞噬时间、吞噬温度、葡萄球菌浓度对外周血白细胞吞噬百分率(PP)和吞噬指数(PI)的影响,并对其进行形态观察。[结果]细菌浓度是影响白细胞吞噬指数的主要因素,白细胞吞噬指数最高的条件为:温度41℃,时间40 min,菌液浓度为5×106个/ml。各因素对白细胞吞噬指数的影响程度为:细菌浓度>时间>温度。[结论]该研究为白细胞吞噬功能的检测提供了一种较为简便、可靠的方法。     

苦参碱对多重耐药表皮葡萄球菌整合子intⅠ和dfrA的影响

为探索苦参碱对多重耐药表皮葡萄球菌(Staphylococcus epidermidis,SE)整合子整合酶(intⅠ)和二氢叶酸还原酶(dfrA)的影响,试验通过纸片扩散法筛选多重耐药菌株、以肉汤稀释法测定苦参碱对SE的最低抑菌浓度(MIC)和最佳逆转条件;通过普通PCR法测定苦参碱对SE整合子intⅠ和dfrA基因碱基序列的影响;通过荧光定量PCR法测定苦参碱对SE整合子intⅠ和dfrA mRNA表达量的影响,结果显示苦参碱亚浓度作用表皮葡萄球菌后对intⅠ、dfrA基因序列无影响;苦参碱对intⅠ mRNA表达量具有抑制作用,而对dfrA mRNA表达量影响不大,结果表明在SE体内苦参碱可从mRNA水平上抑制SE intⅠ的表达量.     

国内外原料乳中金黄色葡萄球菌风险评估研究综述

近年来我国乳品消费量持续增加,乳品质量安全关乎着人们身体健康和生命安全.金黄色葡萄球菌及其肠毒素引起的乳品中毒事件时有发生,逐本求源地针对原料乳开展金黄色葡萄球菌风险评估研究是十分必要的.本文根据国际食品法典委员会拟定的微生物风险评估的原则和指导方针,从危害识别、危害特征描述、暴露评估、风险描述四方面综述了关于原料乳中金黄色葡萄球菌国内外风险评估的研究现状.通过国外成熟的定量风险评估案例,介绍关于原料乳中金黄色葡萄球菌定量风险评估的基本框架,提出未来应加强构建肠毒素预测模型,较为系统的对原料乳从牧场到餐桌进行全程监控和风险评估的建议.