An inhibition enzyme immunoassay for myelin basic protein

An improved Enzyme Immunoassay for Myelin Basic Protein is described. Myelin Basic Protein covalently attached to glass balls, and Myelin Basic Protein in samples compete with each other for binding of a peroxidase conjugated anti Myelin Basic Protein antibody. The peroxidase activity on the balls is then inversely proportional to the amount of Myelin Basic Protein in the sample. A detection limit of 0.6 ng/ml is demonstrated for diluent or spinal fluid. For plasma a dilution step increases this to 1.8 ng/ml. Both the coated balls and the peroxidase conjugate are stable for long periods. The assay requires no expensive equipment. Although the assay appears to be valid for subcellular fractions spinal fluid and plasma, successful detection of Myelin Basic Protection peptides in clinical samples may require careful selection of suitable antisera. The assay would be very suitable for eventual use with an appropriate monoclonal antibody.     

Protein transfer from isoelectric focusing gels: The native blot

The nonelectrophoretic transfer of proteins from thin (0.5 mm) isoelectric focusing gels to nitrocellulose was complete in 1 h. Blotting was bidirectional with 60% of the protein transferred to the top blot and the remaining 40% to the bottom blot. The use of nondenaturing transfer buffers permits proteins to be blotted in the native state. Immunological determinants are preserved and enzyme activity can be detected on the blots.     

草鱼出血病病毒对其它鱼的感染性研究

用草鱼出血病病鱼分离出的草鱼出血病病毒(Grass carp hemorrhage virus,GCHV)感染其它常见鱼并用ELISA方法检查感染鱼组织提取液,结果表明:青鱼、鲢鱼、布氏鳌条对GCHV抗体呈阳性反应;鲤鱼、鳙鱼、鲫鱼、团头鲂、泥鳅则呈阴性反应。综合感染鱼发病症状及死亡特征,初步认为:青鱼对GCHV是易感的,GCHV能在鲢鱼、布氏蟹条体内增值,但毒力较低,鳙鱼、鲫鱼、团头鲂、鲤鱼、泥鳅能抗GCHV感染。     

The sheep antibody response to repeated infection with Lucilia cuprina.

, , and 1992. The sheep antibody response to repeated infection with Lucilia cuprina. International Journal for Parasitology 22: 1169–1174. The specific serum antibody responses of sheep exposed to 10 consecutive infections of L. cuprina have been analysed by enzyme-linked immuno-sorbent assay and immunoblotting using monoclonal antibodies specific for sheep immunoglobulin isotypes. Recognition of a number of larval excretory-secretory products by IgM antibodies appeared to be non-specific. IgG₁ was the major antibody class stimulated by the infection protocol and marked increases in antibody to specific excretory-secretory antigens were observed. Three molecules of 35, 30 and 25 kDa were particularly recognized although the extent of recognition of these molecules varied considerably between individual sheep serum. A pooled serum composed of sera collected after five to seven infections significantly inhibited larval growth in in vitro cultures when compared to a sera pool consisting of sera collected both prior to infection and after infections 1 and 2. The degree of inhibition was greater when serum with high specific antibody titre was used.     

Settlement of the diazotrophic,phytoeffective bacterial strain Pantoea agglomerans on and within winter wheat: An investigation using ELISA and transmission electron microscopy

The aim of this study was to investigate the ability of Pantoea agglomerans, a plant growth-promoting bacterium, to colonize various regions and tissues of the wheat plant (Triticum aestivum L.) by using different inoculation methods and inoculum concentrations. In addition, the enzyme-linked immunosorbent assay (ELISA) and transmission electron microscopy (TEM) were used to determine: (a) the ability of the bacterial cells to grow and survive both on the surface and within internal tissue of the plant and (b) the response of the plant to bacterial infection. After inoculation, cells of the diazotrophic bacterial strain P. agglomerans were found to be located in roots, stems and leaves. Colony development of bacterial cells was only detected within intercellular spaces of the root and on the root surface. However, single bacterial cells were observed in leaves and stems on the surface of the epidermis, in the vicinity to stomatal cells, within intercellular spaces of the mesophyll and within xylem vessels. Inoculated bacterial cells were found to be able to enter host tissues, to multiply in the plant and to maintain a delicate relationship between endophyte and host. The density of bacterial settlement in the plant in all experiments was about 10⁶ to 10⁷ cells per mL root or shoot sap. Establishment was confirmed by a low coefficient of variation of ELISA means at these concentrations.     

Enzyme-amplified enzyme-linked immunosorbent assay for gibberellin A4 based on the NAD-dependent redox cycle

An antiserum against gibberellin A₄ (GA₄) raised in rabbits and its partially purified antibodies were used to develop radioimmunoassay (RIA) and indirect enzyme-linked immunosorbent assays (ELISAs) for GA₄. Of three immunoassays tested, an ELISA based on the NAD-dependent redox cycle (enzyme-amplified ELISA) had highest sensitivity. Levels of methylated GA₄ detected by this most sensitive method ranged from 0.1 fmol/assay (3.5 fg/assay) to 0.1 pmol/assay (3.5 pg/assay) suggesting applicability of this method to the detection of gibberellins in purified plant extracts.     

Effect of anaesthesia with carbon dioxide on vectoring ability and survival of the planthopper Laodelphax striatellus

This paper describes the effects of carbon dioxide anaesthesia on the ability of the planthopper Laodelphax striatellus to subsequently acquire and transmit maize rough dwarf reovirus (MRDV) and barley yellow striate mosaic rhabdovirus (BYSMV), both propagative in the vector. Anaesthesia reduced the acquisition of MRDV by about 45%, but did not affect acquisition of BYSMV. In serial transfer transmission tests, anaesthesia caused interruption of transmission of both MRDV and BYSMV in about 11% of infectious insects; the effect may be due to impaired ability to find the phloem. The effect was stronger on third instar nymphs than on adults. Transmission of MRDV was resumed 3 days after anaesthesia, but none of the insects that stopped transmitting BYSMV resumed it. The survival of insects which ceased to transmit BYSMV was significantly lower than that of hoppers that continued to transmit. No other effects on survival were detected. A possible interaction of carbon dioxide with BYSMV in L. striatellus is discussed, in the light of effects of carbon dioxide on Drosophila melanogaster infected by some rhabdoviruses.     

Application of enzyme-linked immunosorbent assay for mass examination of strongyloidiasis in okinawa, Japan

The enzyme-linked immunosorbent assay was tested to evaluate whether it could be applicable in screening for mass examination of strongyloidiasis. A total of 2906 inhabitants in three areas (858 in Gushikawa Village, 849 in Nakazato Village and 1199 in Sashiki Town) were screened by the enzymatic assay and approximately 11–30% (11.8% in Gushikawa, 17.0% in Nakazato and 27.7% in Sashiki) were considered to be antibody positive. In the parasitological follow-up examinations of those who were antibody positive, actual infection was found in more than half (51%) the subjects. The overall infection rates estimated from the results reached 5.8% in Gushikawa, 9.1% in Nakazato and 14.0% in Sashiki (mean = 10.4%). The infection rates were significantly higher than those in previous surveys conducted in the same areas. The ELISA technique was found to be useful for strongyloidiasis screening and for seroepidemiological purposes in Okinawa.