免疫层析技术及应用的研究进展

免疫层析技术是一类以纳米级标记物探针作为示踪和标记物的检测技术。此类技术除了具有操作简易、反应迅速和时耗较短等特点外,还兼具低成本和高特异性,尤其适用于临场检测。免疫层析技术的发展随着时代需求也在不断加速和更新,现以经典的胶体金免疫层析技术为基础,综述时间分辨与镧系元素荧光免疫层析、荧光微球免疫层析、量子点免疫层析、纳米模拟酶免疫层析及结合新技术的免疫层析的原理、研究进展和临床应用,并对免疫层析技术的未来发展做出展望。     

板栗中毒死蜱农药残留检测结果满意度的研究

本文就如何保证农产品农药残留能力验证实验结果满意需要注意几个关键点进行探讨分析。要想通过板栗中毒死蜱农残的能力验证,样品前处理采用NY/T 761的方法,必须注意：提取液的过滤要用定性滤纸或脱脂棉,提取液过滤后静置时间控制在30 min～40 min之间,吸取10 mL乙腈液体要准确,净化步骤的水浴温度要严格按照方法要求,氮吹切记不要吹的太干,吹到潮乎乎为宜,SPE小柱的活化平衡要充分,上样时液面不能流干,上样后尽可能流干再洗脱,最后定容5.00 mL要准确。检测结束后,对结果进行验证很重要,可采用盲样的空白基质基质配标和空白基质基质加标画标准曲线两种办法。     

一种快速低廉的PCR探针标记方法

【研究目的】探针标记技术是分子生物学和基因工程研究中常用实验技术,需要发展高效、快速、低成本、安全、简捷的DNA探针标记方法;【方法】利用普通的Taq酶进行PCR探针标记,电泳和Southern杂交检测探针标记结果;【结果】电泳检测发现标记产物在电泳时相对非标记的对照表现明显滞后,表明探针标记成功;电泳条带清晰明亮,表明探针标记效率高;Southern杂交条带清晰,说明该方法标记的探针可用于Southern等分子杂交实验;【结论】利用普通的Taq酶进行PCR探针标记,是一种低成本、高效、快速的DNA探针标记方法。     

IN VITRO EVALUATION OF FELINE LEUKOCYTES RADIOLABELED IN WHOLE BLOOD WITH 99MTC STANNOUS COLLOID

Technetium-99m stannous colloid (^99mTcSnC) has been used to radiolabel human leukocytes to investigate various inflammatory disorders. We investigated the in vitro behavior of feline leukocytes labeled in whole blood with ^99mTcSnC. Heparinized blood samples were collected from healthy cats and divided into control and test aliquots. The latter were labeled with ^99mTcSnC using a standard procedure. Leukocyte viability was determined for each sample using a trypan blue exclusion test. Labeling efficiency was determined for test aliquots. Test aliquots were layered onto Histopaque-1077^® and centrifuged before measurement of radioactivity of the blood components. Leukocytes from radiolabeled and control samples were washed and incubated with opsonized zymosan particles to allow assessment of phagocytic function. Aliquots were taken from radiolabeled feline leukocyte samples at 1, 3, 4, and 7 h postlabelling. After centrifugation of each aliquot, radioactivity of the supernatant and pellet was measured and the labeling retention determined. Leukocyte viability in both radiolabeled and control samples was >98%. The labeling efficiency was 95.2±0.14%. The distribution of radioactivity in feline blood was found to be 3.4±0.18% in plasma, 39.0±0.37% in erythrocytes, and 57.6±0.38% in leukocytes. Labeled feline leukocytes had phagocytic activity of 90.9±0.18% (control 91.3±0.15%). The radiolabeled leukocytes retained 93.4±0.19% of the radioactivity up to 7 h postlabeling. ^99mTcSnC efficiently labeled feline leukocytes with no effect on viability and minimal effect on phagocytic function. The percentage retention of radioactivity by the leukocytes was still high at 7 h postlabeling.     

Variable use of plant- and soil-derived carbon by microorganisms in agricultural soils

In this study we used compound specific ¹³C and ¹⁴C isotopic signatures to determine the degree to which recent plant material and older soil organic matter (SOM) served as carbon substrates for microorganisms in soils. We determined the degree to which plant-derived carbon was used as a substrate by comparison of the ¹³C content of microbial phospholipid fatty acids (PLFA) from soils of two sites that had undergone a vegetation change from C3 to C4 plants in the past 20-30 years. The importance of much older SOM as a substrate was determined by comparison of the radiocarbon content of PLFA from soils of two sites that had different ¹⁴C concentrations of SOM.The ¹³C shift in PLFA from the two sites that had experienced different vegetation history indicated that 40-90% of the PLFA carbon had been fixed since the vegetation change took place. Thus PLFA were more enriched in ¹³C from the new C4 vegetation than it was observed for bulk SOM indicating recent plant material as preferentially used substrate for soil microorganisms. The largest ¹³C shift of PLFA was observed in the soil that had high ¹⁴C concentrations of bulk SOM. These results reinforce that organic carbon in this soil for the most part cycles rapidly. The degree to which SOM is incorporated into microbial PLFA was determined by the difference in ¹⁴C concentration of PLFA derived from two soils one with high ¹⁴C concentrations of bulk SOM and one with low. These results showed that 0-40% of SOM carbon is used as substrate for soil microorganisms. Furthermore a different substrate usage was identified for different microorganisms. Gram-negative bacteria were found to prefer recent plant material as microbial carbon source while Gram-positive bacteria use substantial amounts of SOM carbon. This was indicated by ¹³C as well as ¹⁴C signatures of their PLFA. Our results find evidence to support ‘priming’ in that PLFA indicative of Gram-negative bacteria associated with roots contain both plant- and SOM-derived C. Most interestingly, we find PLFA indicative of archeobacteria (methanothrophs) that may indicate the use of other carbon sources than plant material and SOM to a substantial amount suggesting that inert or slow carbon pools are not essential to explain carbon dynamics in soil.     

青贮玉米-牛粪尿体系的15N标记及氮素转化研究

15N示踪技术已开始应用于畜禽粪便氮素循环与利用研究领域,而15N在畜禽粪便不同组分和不同形态氮素中的丰度与数量将直接影响到畜禽粪便15N示踪去向与氮素实际去向的一致性。为了解15N在畜禽粪便标记过程的转化特点和在标记粪尿的分布特征,本文首先采用改进的、含有15N标记硫酸铵(60 atom%15N)的Hoagland营养液砂培种植15N玉米,然后将15N玉米和普通玉米以55∶45的氮配比作为混合青贮饲料饲喂1头已空腹2 d的2龄黄牛,饲喂4 d后停喂2 d,收集全部牛粪尿并对其不同组分和形态氮素的15N丰度和数量进行分析。结果表明:标记玉米、混合青贮饲料、牛粪尿的15N丰度分别为48.024%、26.579%和8.044%;标记玉米对硫酸铵15N的回收率为26.3%,牛粪尿对标记玉米15N回收率为36.0%。在收集的牛粪尿氮中,牛粪全氮、牛尿全氮、牛粪铵态氮和牛尿铵态氮量分别占70.25%、29.75%、5.44%和0.03%,其15N丰度分别为9.223%、5.261%、6.505%和5.419%。在短期内通过饲喂黄牛15N青贮饲料制备的标记牛粪尿中,15N丰度在不同组分和形态氮素中的分布并不相同,牛尿氮的15N丰度低于牛粪氮,矿质态和易于矿化态氮的15N丰度低于不易矿化态氮。     

梨叶片中苹果褪绿叶斑病毒的引物原位标记检测

 选用带苹果褪绿叶斑病毒的香梨叶片，制备成适合进行原位RT-PCR的石蜡切片。设计特异引物，利用引物原位标记技术对切片组织中的病毒进行了检测研究，结果表明，PRINS-FITC染色法和PRINS-Rhodamine染色法均能获得良好的检测效果，有病毒部位分别显示绿色荧光和红色荧光，而且荧光出现的位置与原位RT-PCR检测的结果一致。由此证明引物原位标记技术可用于果树病毒的原位检测。     

Transformation of Upland Cotton （Gossypium hirsutum L.） with gfp Gene as a Visual Marker

The green-fluorescent protein (gfp) gene was evaluated as a screening marker during cotton (Gossypium hirsutum L.) transforming and plant regeneration.High expression of GFP (green-fluorescent protein)...     

转基因产品的标识管理

随着生物科学技术的迅猛发展,转基因产品已经引起方方面面的关注,从国际大环境以及各国所实行的转基因产品的管理政策和方法入手,陈述了我国施行转基因标识管理的必要性和可能性。     

应用~(109)Cd~(2+) ~(65)Zn~(2+)示踪技术研究玉米P Cd Zn间的交互作用

模拟土壤环境,采用109Cd2 和65Zn2 核素双标记示踪技术,以玉米为指示植物研究了P、Cd、Zn间的交互作用,P的浓度设计为0.25(CK)、0.6、3.0mmol·L-1,Cd的浓度为6.5×10-3mmol·L-1;Zn的浓度为0.1mmol·L-1。研究结果表明,在P浓度为0.25和0.6mmol·L-1时,植物根系和地上部对109Cd和65Zn的吸收活度都呈增加趋势,当P浓度为3.0mmol·L-1时,植物根系和地上部对109Cd和65Zn的吸收活度下降,说明P的浓度达到一定水平时,会使重金属Cd和Zn的有效态降低,从而降低了植物对重金属的吸收量。通过对植物根系和地上部吸收的Cd和Zn的比较,发现大多数的Cd和Zn积累在根系,从根部输送到地上部的很少。一是因为植物根系从溶液中吸收重金属后,P与Cd或Zn形成难溶物沉淀下来,使从根系输送到地上部的有效态Cd和Zn显著减少(P≤0.01);二是因为根细胞壁表面带有较多的羧酸基团及少量带正电荷的氨基,Zn2 和Cd2 通过静电吸引而吸附在细胞壁上,减少了Cd和Zn向地上部的输送。从研究结果还可看出,根系吸附的Cd的量大于Zn的量,而茎叶与此相反,这是因为Cd为植物非必需元素,Zn为植物必需的微量元素,植物会把其所需要的元素优先输送到地上部,供其生长发育,所以大多数Zn通过木质部被输送到地上部,大多数Cd被滞留在根系中。利用统计方法分析了相同P水平条件下玉米不同部位吸收的Cd和Zn之间的相关性,结果表明根系吸收的Cd和Zn之间及茎叶吸收的Cd和Zn之间的相关性显著(P≤0.01),Cd与Zn之间存在着协同效应。