Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
1996-08-02
2002-08-27
Minnifield, Nita (Department: 1645)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S388100, C530S388200, C530S388250, C530S389100, C530S389200, C530S389300, C530S391100, C530S391300, C530S413000
Reexamination Certificate
active
06441139
ABSTRACT:
FIELD OF THE INVENTION
The present invention relates generally to an immunologically reactive molecule (IRM) capable of binding to a target molecule from at least one species of insect but not from at least one other species of insect. The present invention also contemplates a method of distinguishing between, or identifying, one or more insect species or subspecies.
BACKGROUND OF THE INVENTION
Insect damage to crops is a significant factor in decreasing crop yield and considerable effort is expended annually in controlling insect populations to minimise or prevent crop damage. Although chemical insecticides have played an important role in controlling insect populations, increasing resistance to these insecticides and environmental concerns have rendered the use of chemical insecticides less appealing.
Resistance Management Strategies (RMS's) have been developed in order to minimize insecticide resistance and usually involve the administration of more than one insecticide. However, such strategies are significantly more expensive, are time consuming and do not address the environmental concerns.
The insect genera Heliothis and Helicoverpa comprise economically important pests which attack a range of crops including cotton, tobacco, maize, sorghum, sunflower, soybean, chick peas and other pulses, rapeseed, canola, ground nuts, lucerne and many horticultural crops such as cut flowers and tomatoes and other vegetables (Fitt, 1989).
Helicoverpa
(Heliothis)
armigera
and
Helicoverpa
(Heliothis)
punctigera
(Lepidoptera: Noctuidae) (hereinafter referred to as
H. armigera
and
H. punctigera
) cause crop damage which is economically significant to agricultural industry. Whilst
H. punctigera
is the more abundant species in Australia,
H. armigera
is considered the more problematic of the two because it has a history of developing resistance to the chemical insecticides which have been used in its control (Fitt, 1989).
Helicoverpa punctigera,
however, does not appear to have the same propensity for developing resistance to insecticides.
Heliothis virescens
and
Helicoverpa zea
(hereinafter referred to as
H. zea
) are important pests of crops throughout North America and especially crops in the U.S.A. Whilst both species have developed resistance to some insecticides, (Fitt, 1989) they are distinguished by the fact that
Heliothis virescens
may display up to 150 fold resistance to synthetic pyrethroids whereas
H. zea
shows no detectable resistance to these compounds (Elzen et al., 1992). Synthetic pyrethroids are therefore more efficacious for control of
H. zea
than for
Heliothis virescens.
Heliothis virescens
is congeneric with
Heliothis
(Neocleptria)
punctifera
whereas
H. zea
is congeneric with
Heliocoverpa armigera
and
Helicoverpa punctigera.
Resistance Management Strategies developed for
H. armigera
and
H. punctigera
have involved a rotation of pyrethroids, endosulfan and other insecticides to reduce exposure to endosulfan and pyrethroids. This has been necessary due to the increasing resistance to synthetic pyrethroids and endosulfan. Despite these RMS's, the frequency of resistant individuals has been steadily increasing (Forrester, 1990).
Helicoverpa armigera
and
H. punctigera
at their egg and neonate stages are morphologically indistinguishable, whilst their respective larvae can only be distinguished by experts. A similar situation exists for
Heliothis virescens
and
H. zea.
Current RMS therefore make assumptions based on historical records of the relative abundance of the two species, averaged over wide geographical distances and several seasons. These assumptions necessarily entail some degree of inaccuracy, the consequence of which is that, in some cases, expensive alternative insecticides may be applied to
H. punctigera,
whilst in other cases, there may be multiple applications of pyrethroids to populations of
H. armigera
. In the latter case, the pyrethroid applications may be ineffective and will generally encourage the emergence of insecticide resistance. Similar problems also arise with
Heliothis virescens
and
H. zea.
Such situations may lead to unnecessarily expensive insect control for farmers. For example, insecticides such as pyrethroids which are adequate for species without a propensity to develop resistance are comparatively cheap insecticides. These may cost the farmer 3 or 4 times less to apply than those insecticides which are applied to insect species with a tendency to develop insecticide resistance. There is a need, therefore, to develop a convenient method to determine the species composition of egg lays, neonates or older larva which may have survived an insecticide application in the field. Such a method would be particularly useful for routine use by crop growers/farmers, agronomists and/or agricultural scientists so that insecticide applications can then be tailored to the composition of the Heliothis or Helicoverpa population actually present.
In work leading up to the present invention, the present inventors have found an immunologically reactive molecule (IRM) that distinguishes between insect species or subspecies. The IRM is capable of binding to a target molecule in at least one species or subspecies but does not react with at least one other species or subspecies. This provides a basis for distinguishing between or identifying insect species or subspecies.
SUMMARY OF THE INVENTION
Accordingly, one aspect of the invention relates to an immunologically reactive molecule (IRM) capable of binding to a target molecule from at least one species or subspecies of insect but not to at least one other species or subspecies of insect. The target molecule is an antigen which is present in insect tissues. These tissues may be those of eggs, neonates or older larvae.
Another aspect of the present invention contemplates a method of identifying one or more species or subspecies of insect by determining the reactivity of an IRM to a target molecule in a sample of insect material, wherein said IRM is capable of binding to the target molecule present in at least one of said species or subspecies but substantially not to any molecule of at least one other species or subspecies of insect. More particularly, the method comprises contacting a sample of insect material with an effective amount of an IRM specific for said target molecule of the one or more species or subspecies of insect for a time and under conditions sufficient for a complex to form between the target molecule and IRM, and then subjecting said complex to a detecting means.
Another aspect of the present invention relates to a kit for distinguishing between or identifying one or more insect species or subspecies from at least one other insect species or subspecies, said kit comprising in compartmentalised form a first compartment adapted to receive an IRM capable of binding to a target molecule of at least one species or subspecies of insect but not at least one other species or subspecies of insect and at least one other compartment adapted to contain a detector means.
In another aspect the instant invention relates to an isolated or purified preparation of a target molecule as herein described specific for a particular insect species or subspecies, to mutants and derivatives thereof and to the antigenic determinants of the target molecule.
In another aspect of the invention there is provided a method for making antibodies, particularly monoclonal antibodies, to said antigenic determinants for use in a method of detecting particular insect species. Antibodies can be made by standard techniques such as immunizing animals with insect extracts from said at least one species or subspecies and collecting the serum, identifying antibodies from said serum which are capable of binding at least one species or subspecies but not at least one other species or subspecies (Campbell (1984), Goding (1983) and Harlow and Lane (1988)).
DETAILED DESCRIPTION OF THE INVENTION
The present invention is predicated in part on the surprising discovery that different
Garsia Kim Astrid
Hill Amanda Susan
Lang Gabriele Annerose
Skerritt John Howard
Trowell Stephen Charles
Brezner David J.
Commonwealth Scientific and Industrial Research
Dorsey & Whitney LLP
Kosslak Renee M.
Minnifield Nita
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