Evaluation of Coagglutination Test Kit for Red Sea Bream Iridovirus

Coagulation test, in principle, is an immunodiagnostics technique, in which immunoglobuline G of antibody is bound to protein A from Staphylococcus aureus. The aim of study is to develop a rapid test kit for detecting iridovirus infection in fish. Method was summarized as follows： （1） vaccine of iridovirus was injected to rabbit four times with a dosage as 0.5 mL, 1 mL, 2mL, 3 mL each week. Serum was collected at the fifth week as a coagglutination test kit; （2） through the positif polymerase chain reaction （PCR） test, the kidney and spleen sample infected with iridovirus are homogenized by using the phosphate buffered saline （PBS） solution of pH 7.2 with ratio 1：2 （WN）; （3） the supernatant material is collected after centrifugation at 8,000 rpm for 15 min; （4） filtrate/supernatant from sample was dropped on a slide an added with coagglutination test kit with the same volume （l：l）; （5） the agglutination observation is done after the 30, 60 and 90 min incubate at room temperature. The coagglutination test gave positive result in 25% of the test samples.

Modification of colorimetric method based digital soil test kit for determination of macronutrients in oil palm plantation

It is the need of time that oil palm farmers must perform the spatially planned soil analysis to know about the fertilizer sufficient and deficient zones of land.Colorimetric method is a suitable and fast solution of soil analysis for NPK determination using the digital soil test kit.NPK determination procedure with a digital soil test kit was undefined for oil palm.Furthermore,the digital soil test kit determines the passage of light through an opaque medium of soil solution with a specified reagent.Therefore,environmental light may interfere leading to wrong results of NPK measurement.Likewise,this equipment was non-incorporable with the controller of any VRT fertilizer applicator.In this research,these issues were addressed and the NPK measurement procedure was defined for oil palm plantation by modifying the‘soil to water’ratio in sample soil solution with an optimum environmental light range of 18-23 W/m^(2).‘Soil to water’ratios were found for nitrogen,phosphorus and potassium as 0.31 to 5.00,1.00 to 5.00 and 4.50 to 5.00,respectively to fit the requirement of NPK for oil palm in the prescribed range of the equipment.Validation study of modified digital soil test kit showed that 91.7%N,89.6%P and 93.8%K results of modified digital soil test kit were matched with analytical laboratory method.Thus,the reliability of NPK results using digital soil test kit was enhanced,making the kit incorporable with the controller of variable rate fertilizer applicator through remote monitoring based data acquisition system.The outcome of this research can be used in the development of an IoT network data fusion for dynamic assessment of the NPK variation in the soil and nutrient management in oil palm plantations.

Development and Performance Measurement of Rapid Detection ciELISA Kit for Ractopamine

Mixed anhydride(MA)was used to conjugate ractopamine(RAC)to BSA and obtained artificial antigen BSA-RAC identified by UV and SDS-PAGE.Balb/c mice were immunized with BSA-RAC and hybridoma lines that secrete RAC monoclonal antibody(mAb)were generated with cell fusion.A ciELISA kit for detection of RAC(RAC-Kit)was developed with RAC mAb and its performance were tested.The results indicated that BSA-RAC was successfully synthesized and its conjugation ratio of RAC to BSA was about 24.5∶1.Three hybridoma lines were filtered and the best one was 4D8-3E11,its affinity constant(Ka)was 1.65×1010 L/mol.The limit of detection of RAC-Kit was 0.5 ng/ml and its detection range was 0.5-184 ng/ml.The mean recoveries of RAC spiked in feed were 85.6% and in swine urine were 88.6%.The precision and accuracy of the assay as determined by inter-assay and intra-assay coefficient variation were below 15%.It had 9.4% cross-reactivity(CR%)to dobutamine and little or no CR to other compounds.The validity of RAC-Kit in 4 ℃ was in 180 d.

Detection of Helicobacter pylori:A faster urease test can save resources

AIM: To investigate whether differences in the rapidity of a positive result for Helicobacter pylori can save res ources, by comparing two commercially available urease kits. METHODS: One hundred and eighty-five adults (130 outpatients, 55 inpatients) undergoing gastroscopy were entered prospectively. Patients were divided into two groups: Group 1 (if they were not on PPIs, antibiotics, H2A, bismuth or sucralfate for up to 14 d prior to the endoscopy) and Group 2 (if they were on, or had been on, any of the above medication in the previous 14 d). At endoscopy two sets of biopsies, taken in random order, were placed in the wells of the Campylobacter-like organism (CLO) test (Kimberly-Clark, Utah, USA) and the Quick test (Biohit Plc, Helsinki, Finland). Five additional gastric biopsies were taken for histology/Giemsa and immunohistochemical study. The two urease test slides were read at 2 min, 30 min, 2 h and 24 h. Sensitivity and specif icity at 24 h were determined. RESULTS: At 24 h, for all patients, there was no difference in sensitivity (100% vs 97.5%), specificity (99.3%), positive (97.5%) and negative predictive values (100% vs 99.3%) between the CLO and Quick tests, respectively. There was a positive result at 30 min in 17/41 (41.5%) CLO tests, and in 28/40 (70%) Quick tests, P = 0.05. Quick test enabled the prescription of eradication therapy before discharge in all 28/40 patients. Only 12 (30%) follow-up appointments were needed. If the CLO test had been used alone, only 17 (41.5%) prescriptions would have been possible prior to discharge and 24 (58%) follow-up appointments would be needed (P = 0.001). Of 2000 gastroscopies performed annually at our unit, a saving of 123 follow-up appointments (total: 8856 Euros or 11 808 USD) would be achieved if we switched to the Quick test. CONCLUSION: Direct comparison of locally available urease test kits is worthwhile, since the appropriate choice results in a significant saving of resources. Local costs and follow-up protocols will determine the magnitude of these savings.

保存温度及保存时间对AmpFlSTR~ Identifiler~ Plus Kit PCR Reagents试剂盒PCR混合液效能影响的研究

目的探讨AmpFlSTRIdentifiler Plus Kit PCR Reagents混合液使用时间和温度的影响因素。方法试剂盒PCR混合液依次在4℃、室温、-20℃下保存1周、2周至15周不等,用于样品STR分型检验。结果4℃保存的PCR混合液在15周之内可以检验出标准品9947A的STR分型;室温保存在6周之内可以检出;-20℃保存至少在15周内可以检出。结论由AmpFlSTR Identifiler Plus Kit PCR Reagents试剂盒各组分组成的试剂盒PCR混合液在一定的保存条件下使用是可行的,符合法庭科学人类荧光标记STR技术要求。

基于方差分析研究不同微生物方法对奶粉中泛酸含量测定的影响

采用国标方法、微生物试剂盒法和水浴提取国标法分别对质控样品及奶粉样品进行泛酸含量的测定,利用SPSS统计分析方法对结果进行分析评价。结果表明国标法标准曲线相关系数为R^(2)=0.9982,试剂盒法相关系数为R^(2)=0.9988,两者方法灵敏度高;3种方法的RSD为2.10%~3.11%,均小于5%;使用SPSS 2因素完全随机方差分析法、2因素重复测量分析法分别对质控样品及奶粉样品测定结果进行分析,发现3种不同方法对泛酸含量测定无显著影响(P>0.05)。人员比对实验数据精密度、正确度高、人员操作水平一致,数据显示实验室检验误差小,符合方法要求。结果认为3种方法测定结果并无统计学差异,以此依据对国标方法进行优化,提高了前处理效率,实验室可根据实际需求选用最佳方案。

胆固醇-3,4-^(13)C_(2)的合成

针对我国胆固醇-3,4-^(13)C_(2)产品制备方法空白,以乙酸苯酯-1,2-^(13)C_(2)和4-胆甾烯-3-酮为起始原料,合成胆固醇-3,4-^(13)C_(2)。总体标记合成路线采用胆固醇基础结构拆分后引入标记砌块再重构目标化合物结构的方案,其中4-胆甾烯-3-酮通过开环氧化、合环得到标记前体,乙酸苯酯-1,2-^(13)C_(2)作为^(13)C标记引入砌块与标记前体通过乙酰化反应得到乙酰化标记中间体,最后乙酰化标记中间体再经过碳环重构、酯化、还原反应得到^(13)C二标记的类固醇激素类化合物胆固醇-3,4-^(13)C_(2)。其中还原反应显示出一定的不对称合成现象。以投入的乙酸苯酯-1,2-^(13)C_(2)的物质的量计算反应总产率为16.2%。所得终产品经MS、NMR和HPLC表征确认,液相色谱纯度≥96%,液相质谱检测丰度≥98 atom%^(13)C,该产品可用于临床质谱、生物医药等领域检测用稳定同位素内标试剂以及临床质谱用疾病筛查试剂盒原料。

和光溢彩——铜纳米簇的光致发光现象

本实验自制谷胱甘肽与半胱氨酸分别配位的铜纳米簇(CuNCs),根据CuNCs光致发光原理,设计实验探究配体、氧化剂、pH、溶剂及温度对CuNCs光致发光的影响,并开发出科普实验试剂盒。本试剂盒携带方便、操作简单、安全性高,适合多种科普场景,能够满足具有不同梯度科普需求的人群,增强公众对纳米材料的了解,让公众感受到化学之美。

体外检测试剂盒阻断剂的制备方法及特性分析

目的探讨体外检测试剂盒阻断剂的制备方法及特性。方法采用6周龄BALB/c小鼠进行抗原免疫,构建杂交瘤细胞并克隆。采用动物体内诱导单克隆抗体的方法大量制备单克隆抗体,分别采用G柱亲和层析、蓝胶+Q柱离子交换层析两种方法进行纯化,核酸蛋白检测仪记录波长280 nm下的光密度(optical density,OD),紫外分光光度计测量成品浓度。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecylsulfate-polyacrylamide gel electrophoresis,SDS-PAGE)检测抗体纯度。采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)双抗体夹心法检测该阻断剂(CN302)对假阳性样本的阻断能力。结果G柱亲和层析法提取阻断剂的收率(3个批次分别为6.49、6.12、6.27 g/L)高于蓝胶+Q柱离子交换层析法(3个批次分别为1.78、1.68、1.61 g/L)。两种纯化方法的纯度均高达95%以上。G柱亲和层析法、蓝胶+Q柱离子交换层析法所得阻断剂的活性分别为106.54%、92.45%;与空白组相比,两种纯化方式所得阻断剂CN302对假阳性样本的检出率均降低,其OD值均小于0.2,与基准阻断剂大致相同。G柱亲和层析法所得阻断剂CN302未冻融的活性为99.86%,冻融5次后活性为99.83%。无阻断剂存在时,对假阳性样本检测的OD值约为2.5,最高可达到4.0;加入阻断剂后,OD值明显降低;阻断剂CN302对假阳性样本检测的OD值低于其他阻断剂。结论本研究所得阻断剂能显著消除样本内源性干扰,且活性几乎全部保留。G柱亲和层析法提取抗体的收率和阻断能力均高于蓝胶+Q柱离子交换层析法。

基于超小金纳米颗粒的急性心梗快速检测试剂盒研制

以油胺为还原剂,十八烯和聚乙烯吡咯烷酮为分散剂,借助微流控技术合成了超小球形金纳米颗粒。通过紫外分光光度计(UV-Vis)、X-射线衍射(XRD)、能谱仪(EDX)、高分辨透射电子显微镜(HR-TEM)以及选区电子衍射(SAED)等分析,对超小金纳米颗粒的相结构、成分和形貌进行了全面表征。基于这些超小金纳米颗粒,通过工艺优化,成功制备出了急性心梗快速检测试剂盒,并对其性能进行考察。结果表明:合成的金纳米颗粒具有多晶结构,平均粒径仅4.57 nm;基于上述金纳米颗粒,获取的急性心梗快速检测试剂盒可对心梗进行快速检测,对心梗标志物(心肌肌球蛋白结合蛋白C)的检测限可低至0.1 ng/mL;同时,该试剂盒呈现出了高的特异性、重复性和稳定性,具有应用潜质。

量子点标记的新型冠状病毒抗体试剂盒行业分析

新型冠状病毒感染全世界大流行阶段,随着对新型冠状病毒感染的预防、确诊及治疗等方面认识的不断深入,各种体外检测技术与应用产品成为研发人员关注的重点。相较于传统的基因测序及应用广泛的核酸检测,或同为免疫学检测试剂盒的胶体金免疫层析、酶联免疫吸附法(ELISA)、化学发光等技术,新型纳米材料免疫标记试剂盒是一种兼具快速、简便和灵敏度、特异度良好等优点的新型冠状病毒检测方法。本文通过比较不同新型冠状病毒检测方法,重点阐述新型纳米材料免疫标记新型冠状病毒检测试剂盒的商业价值及其潜在应用价值。

Research Progress in Formation and Detection Method of Dextran in Sugarcane Production

Dextran in sugarcane production process is formed by Leuconostoc rnesenteroides. The content levels of dextran is related to sugarcane varieties, field condition （planting pattern, temperature, humidity, sunlight, soil, foreign material）, de- gree of injury （refractory cane, harvesting methods）, and can be rapidly and accu- rately measured by Dextran Immunonephelometric Test Kit. The presence of dextran indicates that sucrose has been lost, so sugarcane dextran is a direct and reliable indicator to measure sugarcane freshness and quality.

四环素类药物多残留酶联免疫检测方法

【目的】构建一种四环素类药物多残留酶联免疫检测试剂盒。【方法】将四环素类药物与载体蛋白相连作为抗原免疫动物,获得了抗四环素类药物的抗体,建立了动物性食品中四环素类药物残留的ELISA检测方法,并将该试剂盒的检测性能与进口试剂盒比较,对实际样品的测定结果与高效液相-串联质谱法比较。【结果】人工抗原中四环素与蛋白质分子的结合比约为15﹕1,血清效价达到1﹕2000倍以上,50%抑制浓度(IC50)为3μg·L-1左右,7种四环素类药物交叉反应率在58%～100%之间,牛奶和鸡肉中四环素类药物的检测限分别为6.6μg.L-1和6.5μg·kg-1,在鸡肉中四环素、土霉素和金霉素的回收率在40%～120%之间。【结论】试剂盒的最低检测限为0.3μg·L-1,检测范围为0.3～30μg·L-1,可同时测定兽医常用的四环素、金霉素、土霉素和多西环素4种药物残留。灵敏度、精密度和准确度均能满足兽药残留检测要求。

禽流感间接ELISA诊断试剂盒的研制及应用

将成熟的禽流感间接ELISA快速诊断技术试剂盒化 ,确定其外包装、规格、盒内组成 ,对盒内各组分的性状、保存、质量控制以及诊断试剂盒的特异性、敏感性、可重复性、符合率、与国外同类产品比较、保存期等进行了全面、详细的研究。试剂盒由 9种成套试剂组成 ,在 - 15℃至 - 2 0℃保存 5 40天各项性能仍然很好。该试剂盒与进口禽流感间接ELISA诊断试剂盒对同样血清样品检测 ,符合率为 10 0 %。拥有该试剂盒 ,只需准备生理盐水 ,即可对大量血清样品同时进行检测 ,操作简单方便、结果特异性强、敏感性高、稳定、快速 ,更适合于禽流感免疫抗体的监测及现地疫病诊断等需要。先后制备诊断试剂盒 16个批次 ,已成功应用于我国省、市兽医站、兽医研究所、农业院校、农科院、农业公司、口岸检疫部门及鸡场等各领域对禽流感的定性诊断、流行病学调查、免疫抗体监测等。

鸡传染性法氏囊病间接ELISA诊断试剂盒的研制及初步应用

采用 vero细胞增殖的 IBDV抗原建立了间接 EL ISA,用于定量检测 IBDV抗体。对该方法进行了标准化研究 ,确立了该方法的最佳工作条件。在此基础上研制了相应的诊断试剂盒 ,并对盒内各组分的性状、保存、质量控制以及诊断试剂盒的特异性、敏感性、可重复性、符合率、与国外同类产品的比较、保存期等进行了全面、详细的研究。结果表明 ,研制的试剂盒在 - 2 0℃保存至 6个月时各项性能都很好。该试剂盒与进口试剂盒对同样血清样品检测 ,符合率为86 .7%。间接 EL ISA试剂盒与琼扩试验的符合率为 92 .5 %。该试剂盒可对大量血清样品进行检测 ,操作简单方便、结果特异性强、敏感性高、快速 ,更适合于大型鸡场 IBD免疫抗体水平的监测及现地疫病诊断等需要。

链霉素残留检测ELISA试剂盒的研制及应用

本研究采用EDC法人工合成链霉素抗原（SM—BSA），免疫BALB／c小鼠，通过杂交瘤技术获得了分泌抗链霉素单克隆抗体的杂交瘤细胞株，建立了快速检测链霉素（SM）残留的酶联免疫方法，并成功研制了试剂盒。然后对其灵敏度、准确度、特异性、基质效应性等技术指标进行检测，同时应用该试剂盒对生长猪尿液中的残留情况进行检测。结果表明：除双氢链霉素外，该试剂盒与其它抗生素类药物均无交叉反应；线性检测范围为1～1289g／L，相关系数R^2=0．9251，灵敏度为0．45μg／L，半数抑制浓度（IC50）为7．769g／L，检测限为19g／L；猪尿样的平均添加回收率为84．1％；基质对该试剂盒的检测结果影响不大。该试剂盒具有快速、敏感、特异、简便等特点，适合于SM残留的快速检测，具有较高的推广应用价值。

Prev-DAF试剂盒分析线粒体基因1555A-G突变

目的建立应用标准试剂盒方法检测线粒体基因1555A-G(mtDNA1555A-G)突变的程序,进行母系遗传耳聋家系的基因型分析。方法采用Prev-DAF试剂盒分析14个母系遗传耳聋家系,共检测耳聋患者34个,正常个体11个,并以Alw26酶切和测序方法验证试剂盒检测的准确性。结果Prev-DAF试剂盒检测结果证实13个家系中33个耳聋患者携带有mtDNA1555A-G突变,另1个耳聋家系中的一名耳聋患者不携带此突变,11个正常个体中无此突变,试剂盒检测方法与Alw26I酶切法和测序结果完全吻合。结论在中国,mtDNA1555A-G氨基糖甙类抗生素致聋的家系多,分布广,Prev-DAF试剂盒在分析线粒体基因1555A-G突变方面具有简单、低耗、结果直观的特点,适合在中国用于进行此突变的大规模筛查或预防性检查。

氯霉素残留检测阻断ELISA试剂盒的研制及性能测定

在建立氯霉素单克隆抗体(CAP mAb)杂交瘤细胞株和阻断ELISA方法的基础上,研制出CAP残留快速检测试剂盒(CAP-kit),并对其性能进行了测定。结果表明,CAP-kit的标准曲线呈典型的S型,符合4参数logit曲线拟合,相关系数R2=0.9953,检测范围为1.0μg/L～128.0μg/L,灵敏度为0.85μg/L,半数抑制浓度(IC50)为7.54μg/L,检测限为1.0μg/L;牛奶样、猪肉样的平均添加回收率为88.3%、82.5%,平均批内和批间变异系数均<15%;CAP-kit与氯霉素琥珀酸钠的交叉反应(CR%)为150%,与其它酰胺醇类和抗菌素药物无交叉反应;试剂盒在4℃可保存6个月。

猪圆环病毒2型ELISA抗体检测试剂盒的研制及应用

利用纯化的重组杆状病毒表达猪圆环病毒2型(PCV2)Cap蛋白作为检测抗原,建立一种间接ELISA方法用于血清抗体检测。昆虫细胞株(Sf-21)接种重组杆状病毒,对蛋白表达参数及纯化工艺进行了优化,制备了5批重组蛋白抗原,表达量占总蛋白的18.7%左右;Ni2+亲和层析柱纯化获得重组蛋白纯度为90.1%;重组蛋白可被特异性抗体识别,具有良好的免疫活性。用建立的ELISA法对30份已知阴性血清样本检测临界OD405nm值为0.343;ELISA与IPMA对150份血清进行平行检测符合率为95.3%,特异性为91.2%,敏感性为96.5%;与其他几种常见猪病毒参考血清无交叉反应。组装的试剂盒批内和批间变异系数分别为4.0%～4.8%和8.9%～9.9%;置-20℃保存12个月稳定。试剂盒对人工感染猪血清检测结果,接种后第5周抗体阳转,第8周～9周抗体水平达到高峰;对来自黑龙江、吉林、河北、上海等地猪场1085份血清抗体检测,阳性率为73.6%。研制的ELISA试剂盒为临床PCV2血清抗体检测及疫苗免疫效果的评价提供了技术手段。

莱克多巴胺单克隆抗体的研制及阻断ELISA检测方法的建立

用混合酸酐法将莱克多巴胺（RAC）偶联于牛血清白蛋白（BSA）,合成免疫原BSA-RAC,并用紫外和凝胶电泳鉴定;用BSA-RAC免疫Balb/C小鼠,细胞融合技术建立抗RAC单克隆抗体（mAb）杂交瘤细胞株,体内诱生腹水法制备RAC mAb。应用RAC mAb研制RAC残留快速检测阻断ELISA试剂盒（RAC-Kit）,并测定其性能。结果表明,BSA-RAC偶联成功,分子结合比为1∶24.5;筛选出3株杂交瘤细胞,其中最好的4D8株的亲和常数Ka为1.65×1010L/mol。RAC-Kit的标准曲线呈典型的S型,符合4参数logit曲线拟合,线性范围为1~170μg/L,灵敏度为0.52μg/L,检测限为1μg/L,饲料样、猪尿样的平均添加回收率分别为91.1%和89.2%,平均批内和批间变异系数〈15%,与多巴酚丁胺的交叉反应率为22.3%,与其他化合物无交叉反应,RAC-Kit在4℃可保存180d。