Method for detecting antigens and antibodies

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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Details

435 12, 435188, 435810, G01N 3350, C12Q 158, C12N 996

Patent

active

045901570

DESCRIPTION:

BRIEF SUMMARY
This invention relates to improvements in methods, test kits and apparatus for performing enzyme-linked immunosorbent assays for the detection and quantitative determination of antigens and antibodies. In particular, the present invention relates to improvements in methods, test kits and apparatus which enable such assays to be performed by following a simple procedure which does not require the use of elaborate laboratory equipment or trained laboratory personnel, and hence may be performed under relatively adverse conditions, for example in the field.
In prior Australian Patent Specification No. 68117/81, there are described improved methods and apparatus for the performance of enzyme-linked immunosorbent assays (ELISA), which are particularly described with reference to their use in the detection of anti-tetanus antibodies in sheep and the detection and identification of snake venoms as examples of such assays.
The prior specification describes apparatus for detecting or determining the presence of antigenic or haptenic substances or antibodies in a sample which comprises a plurality of containers, one of said containers being provided with a cap which has antibodies or antigenic or haptenic substances attached to an internal surface thereof, said cap being adapted to be transferred from one to another of said containers so the internal surface thereof is brought into contact with solutions contained in said plurality of containers in a predetermined sequence. In one embodiment the "internal surface" of the cap includes those surfaces of the cap themselves which, when the cap is applied to a container, are exposed to the contents of the container, as well as the surfaces of projecting portions provided on the cap to extend within the body of the container when the cap is applied thereto, and surfaces which are physically attached to or retained within a cap which are themselves exposed to the contents of the container when the cap is applied thereto. In an alternative embodiment, the "internal surface" of the cap to which the antibodies or antigenic or haptenic substances are attached comprises the internal surface of a tube attached to the cap, for example the tube portion of a "dropper-type" cap having a rubber teat or the like to enable the fluid to be drawn up into the tube and expelled therefrom.
One of the main limitations of the apparatus disclosed in the prior specification lies in the fact that only a single antibody or antigenic or haptenic substance can be attached to the internal surface of the cap or of the tube attached to the cap. This is a relatively serious limitation when the apparatus is to be used in a screening-type test. For example, in the detection and identification of snake venoms in Australia, it is usually necessary to be able to detect at least two and in most cases five primary snake venoms to enable selection of the appropriate antivenom. This means that when using the apparatus disclosed in the prior specification, at least two and often five separate tests must be carried out in order to identify the unknown venom in the clinical sample, together with control tests where appropriate. The necessity to carry out a number of separate tests is, of course, particularly disadvantageous from the point of view of carrying out the necessary tests in the field, and it is an object of the present invention to provide a further improvement in this apparatus whereby the requirement for performing a plurality of separate tests may be avoided.
According to a first aspect of the present invention there is provided a device for use in detecting or determining the presence of antigenic or haptenic substances or antibodies in a sample, which device comprises a plurality of tubular or capillary elements, each of said capillary elements having antibodies or antigenic or haptenic substances attached to an internal surface thereof, and means for causing fluids to pass simultaneously or sequentially through said plurality of capillary elements.
In use of the device of this aspect of the inventi

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patent: 4299916 (1981-11-01), Litman et al.

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