WO2020232313A1

WO2020232313A1 - Lateral flow device for target analyte detection using chemosensory proteins

Info

Publication number: WO2020232313A1
Application number: PCT/US2020/033003
Authority: WO
Inventors: Daniel F. Woods; Ken Konrad
Original assignee: Inscent, Inc.
Priority date: 2019-05-14
Filing date: 2020-05-14
Publication date: 2020-11-19

Abstract

The present specification discloses a lateral flow device for detecting an analyte, a detection device comprising the disclosed lateral flow device, testing kits and testing containers comprising the disclosed lateral flow device or the disclosed detection device, and methods and uses for detecting an analyte in a sample using the disclosed lateral flow device, disclosed detection device, disclosed testing kits or disclosed testing containers.

Description

LATERAL FLOW DEVICE FOR TARGET ANALYTE DETECTION USING

CHEMOSENSORY PROTEINS

[001] This application claims the benefit of priority and is entitled to the filing date pursuant to 35 U.S.C. § 119(e) of U.S. Provisional Patent Application 62/847,860, filed May 14, 2019, the content of which is hereby incorporated by reference in its entirety.

[002] Ensuring water supplies are safe is crucial to public health. Water supply safety concerns are not limited to potable water or even water that is intended for domestic use; irrigation water and water sources not intended for direct human use or consumption must also meet basic safety and hygiene requirements in order to ensure public safety and prevent disease transmission from water-borne microbes. As one example, an indicator organism that reveals contamination in a tested water sample is the bacterium, E. coli. However, present methods approved by the US Environmental Protection Agency for detecting E. coli contamination require at least 24 hours. Given the relatively short time required to distribute foods such as vegetables that may have been treated with water, a reliable detection mechanism requiring less time would improve water safety. An interesting characteristic shared by many current contamination detection methods is the essentially retroactive or ex post facto nature of their results; however, since point source contamination of water supplies is sometimes swift, a quick, field-deployable device would address water safety concerns on a more immediate basis. Deadly outbreaks of E. coli 0157:H7, such as the incidents involving spinach grown in California and lettuce grown in Arizona, highlight the need for a rapid, effective and cost-efficient detection methods, for E. coli as well as other pathogens, as modem distribution networks can quickly result in contaminated food infections spanning large geographic areas. For example, the 2018 E. coli 0157:H7 outbreak from Arizona lettuce resulted in 210 cases of infection in 36 states, including 96 hospitalizations and 5 deaths; one death involved a patient infected across the country in New York. Accordingly, a field-deployable, easy to use biosensor capable of providing rapid results in response to point- source contamination of the water supply would have been particularly valuable in these instances.

[003] Insects rely on several classes of chemosensory proteins to detect scents and tastes from the environment and to regulate crucial responses to environmental stimuli. The odorant-binding proteins (OBPs) are the first chemosensory proteins to bind to an odor or scent molecule from the environment, and are thus responsible for the initial step of molecule recognition. A highly successful and diverse group, insects have evolved a large variety of OBPs that are capable of binding to numerous molecules. Insect OBPs are relatively small (less than ~20 kDa) proteins assembled with six a-helices and are additionally stabilized by disulfide bridges to yield a robust and resilient structure. As a result, insect OBPs are characterized by thermal stability and resistance to proteolysis and denaturation as well as the capacity to readily refold upon restoration of favorable conditions. Furthermore, many insect species rely on odor detection to regulate crucial behaviors such as feeding and mating, and this detection of food sources and mates is mediated by insect OBPs and the closely related pheromone-binding proteins (PBPs) respectively. Insect OBPs have consequently evolved into a diverse family of proteins with a wide range of analyte specificities and selectivities. These robust, diverse proteins are thus well suited for service as detector elements in novel biosensors.

[004] The advantageous physical and binding properties of insect OBPs have resulted in several attempts at utilizing these proteins in biosensor devices to detect a variety of analytes. OBP14 from the honeybee, Apis mellifera, has been used to detect the neuroblastoma biomarker, homovanillic acid, in a device based on reduced graphene oxide (rGO) field-effect transistor (FET) technology. The ligand specificity and selectivity of recombinantly expressed Apis mellifera OBP14 can be fine-tuned by generating mutant variants; for example, an additional disulfide bridge in the protein’s structure will increase its affinity for a component of plant odors that is also an insect pheromone precursor, eugenol. In fact, the concept of fine-tuning biosensor responses by introducing mutations to an OBP detector element has been explored successfully and may prove useful in future biosensor devices. An rGO-FET OBP 14 biosensor can also be used to detect compounds attractive to bees. Likewise, OBP biosensors relying on electrochemical impedance measurements are arguably feasible and current technologies have made the artificial“electronic nose” realistic. These and similar insect OBP-based biosensors demonstrate that proteins derived from the insect chemosensory system are suitable for use as detector elements in biosensors; however, these biosensors are generally complex devices with reporter mechanisms that rely on advanced technologies.

[005] Therefore, there is a need for effective, stable, robust devices for target analyte detection, as well as related testing device kits and/or testing containers, as well as related methods of use. These new devices and associated methods of use overcome at least the traditional approaches enumerated above. Accordingly, and in one particular aspect of the disclosure, biosensor implementations based on an insect OBP, AgamOBPl from Anopheles gambiae, as the detector element, are disclosed below. In particular embodiments described below, the biosensors can detect a characteristic bacterial metabolite, such as indole, quickly and with high sensitivity, making the devices suitable as first-line means of detecting coliform bacterial contamination in water supplies. Other devices and methods of use are possible and are provided herein in accordance with the teachings of the present disclosure, as discussed more fully below.

SUMMARY

[006] In the developing world, the identification of clean, potable water continues to pose a pervasive challenge, and waterborne diseases due to fecal contamination of water supplies significantly threaten public health. The ability to efficiently monitor local water supplies is key to water safety, yet no low-cost reliable method exists to detect contamination quickly. In one aspect of the present disclosure, an in vitro assay utilizing an odorant-binding protein (OBP), AgamOBPl, from the mosquito, Anopheles gambiae, to test for the presence of a characteristic metabolite, indole, from harmful coliform bacteria is provided. As disclosed herein, recombinantly expressed AgamOBPl binds indole with high sensitivity. In one aspect of the disclosure, the assay is fluorescence-based and demonstrates the usefulness of insect OBPs as detector elements in novel biosensors that rapidly detect the presence of a target analyte, such as, but not limited to, bacterial metabolic markers, and thus of coliform bacteria. In a particular aspect, rAgamOBPl is herein disclosed to be suitable for use in portable, inexpensive“dipstick” biosensors that improve upon lateral flow technology since insect OBPs are robust, easily obtainable via recombinant expression, and resist detector“fouling.” Moreover, due to their wide diversity and ligand selectivity, insect chemosensory proteins have other biosensor applications for various analytes, as disclosed herein and in accordance with the teachings of the present disclosure. Techniques, devices, methods and kits disclosed herein represent platform technologies applicable to various exemplary devices and method of use, in accordance with the present disclosure, for detecting the absence or presence of target analytes.

BREIF DESCRIPTION OF DRAWINGS

[007] The accompanying Figures illustrate particular aspects of the present invention.

[008] FIG. 1 shows detection of indole using an embodiment of a lateral flow device disclosed herein; [009] FIG. 2 shows detection of E. coli strain K-12 cells using an embodiment of a lateral flow device disclosed herein;

[010] FIG. 3 shows detection of fecal contamination using an embodiment of a lateral flow device disclosed herein;

[Oil] FIG. 4A-D shows schematics of an exemplary lateral flow device disclosed herein with FIG. 4A showing a top perspective view of a lateral flow device with cut-away revealing internal components; FIG. 4B showing top plan view of a lateral flow device illustrating a negative result; FIG. 4C showing top plan view of a lateral flow device illustrating a positive result; FIG. 4D showing a schematic of lateral flow device analyzing sample lacking target analyte (a negative result); and FIG. 4E showing a schematic of lateral flow device analyzing sample containing target analyte (a positive result); and

[012] FIG. 5 shows sensitivity of an exemplary lateral flow device to various dilutions of indole.

DETAILED DESCRIPTION

[013] Now turning to a non-limiting introduction to broad aspects of the present specification, a lateral flow device for detecting the presence or absence of a target analyte in a sample is herein disclosed. In some embodiments, a disclosed lateral flow device comprises a matrix that supports the flow of a liquid sample, such as, e.g., by capillary flow of the fluid. In some embodiments, a disclosed lateral flow device comprises a matrix that supports the flow of a liquid sample and a solid support that provides structural stability to the matrix. As discussed in greater detail below, a matrix can be composed of a single material or a plurality of different materials. Non-limiting examples of a solid support include a laminate backing material comprised of mylar or polyester.

[014] A matrix disclosed herein comprises several zones. In some embodiments, a matrix comprises a competitive reagent zone and a detection zone. In some embodiments, the matrix comprises a competitive reagent zone, a detection zone and an absorbent zone. In some embodiments, the matrix comprises sample application zone, a competitive reagent zone, and a detection zone. In some embodiments, the matrix comprises a sample application zone, a competitive reagent zone, a detection zone and an absorbent zone. The preferred order of these zones, in the direction of capillary flow is a sample application zone, a competitive reagent zone, a detection zone and an absorbent zone.

[015] A competitive reagent zone of a matrix disclosed herein comprises a chemosensory protein conjugate. In some embodiments, a chemosensory protein conjugate includes a chemosensory protein. The particular chemosensory protein selected for a disclosed lateral flow device will depend on the target analyte that the lateral flow device is designed to detect. In some embodiments, a chemosensory protein used in a lateral flow device disclosed herein can bind indole or a derivative of indole. In some embodiments, a chemosensory protein used in a lateral flow device disclosed herein is an odorant binding protein. In some embodiments, a chemosensory protein used in a lateral flow device disclosed herein is an odorant binding protein that can bind indole or a derivative of indole. In aspects of these embodiments, an odorant binding protein includes an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33. In other aspects of these embodiments, an odorant binding protein is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 87%, at least 90%, at least 93%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4. In some embodiments, a chemosensory protein conjugate includes a chemosensory protein disclosed herein covalently linked to a nanoparticle. Non-limiting examples of a nanoparticle include a colloidal gold particle, a colored latex particle, a carbon particle, a selenium particle, a chemiluminescent particle, a bioluminescent particle, a fluorescent particle, a quantum dot particle, an upconverting phosphor particle, a liposome including a dye.

[016] A nanoparticle disclosed herein is generally about 15 nm to about 800 nm in diameter. In aspects of this embodiment, a nanoparticle disclosed herein is about 25 nm to about 700 nm, about 50 nm to about 600 nm, about 75 nm to about 550 nm, about 100 nm to about 500 nm, about 125 nm to about 500 nm, about 150 nm to about 450 nm, about 200 nm to about 400 nm, about 250 nm to about 350 nm or about 300 nm to about 325 nm, for example. In particular embodiments, such a nanoparticle can have a size of about 15 nm, about 30 nm, about 45 nm, about 60 nm, about 75 nm, about 90 nm, about 105 nm, about 120 nm, about 135 nm, about 150 nm, about 165 nm, about 180 nm, about 195 nm, about 205 nm, about 220 nm, about 235 nm, about 250 nm, about 265 nm, about 280 nm, about 295 nm, about 310 nm, about 315 nm, about 330 nm, about 345 nm, about 360 nm, about 375 nm, about 390 nm, about 405 nm, about 420 nm, about 435 nm, about 350 nm, about 365 nm, about 380 nm, about 395 nm, about 405 nm, about 420 nm, about 435 nm, about 450 nm, about 465 nm, about 480 nm, about 495 nm, about 510 nm, about 525 nm, about 540 nm, about 555 nm, about 570 nm, about 585 nm, about 600 nm, about 615 nm, about 630 nm, about 645 nm, about 660 nm, about 675 nm, about 690 nm, about 705 nm, about 720 nm, about 735 nm, about 750 nm, about 765 nm, about 780 nm, about 795 nm or about 800 nm, for example.

[017] A detection zone of a matrix disclosed herein comprises a plurality of capture reagent zones. In some embodiments, a detection zone comprises a first capture reagent zone and a second capture reagent zone. A first capture reagent zone comprises a competitive ligand to the chemosensory protein comprising the chemosensory protein conjugate of the competitive reagent zone. A first capture reagent zone serves as an indicator for the presence or absence of the target analyte. In some embodiments, a competitive ligand is one that that binds to a chemosensory protein that can bind indole or a derivative of indole. In some embodiments, a competitive ligand is one that that binds to an odorant binding protein. In some embodiments, a competitive ligand is one that that binds to an odorant binding protein that can bind indole or a derivative of indole. In some embodiments, a competitive ligand is one that that binds to an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33. In other aspects of these embodiments, a competitive ligand is one that that binds to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 87%, at least 90%, at least 93%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4. In some embodiments, a competitive ligand is indole, 3-methyl indole or another derivative of indole. In some embodiments, a competitive ligand is covalently linked directly to the matrix. In some embodiments, a competitive ligand is indirectly attached to a matrix, such as, e.g., a competitive ligand can be covalently linked to a carrier protein which in turn is directly attached to the matrix. Non-limiting examples of a carrier protein include a serum albumin or a thyroglobin.

[018] A second capture reagent zone comprises an antibody having specificity for the chemosensory protein comprising the chemosensory protein conjugate of the competitive reagent zone. A second capture reagent zone serves as a positive control for the chemosensory protein conjugate. In some embodiments, an antibody disclosed herein is an antibody having specificity for a chemosensory protein that can bind indole or a derivative of indole. In some embodiments, an antibody disclosed herein is an antibody having specificity for an odorant binding protein. In some embodiments, an antibody disclosed herein is an antibody having specificity for an odorant binding protein that can bind indole or a derivative of indole. In aspects of these embodiments, an antibody disclosed herein is an antibody having specificity for an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33. In other aspects of these embodiments, an antibody disclosed herein is an antibody having specificity for SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 87%, at least 90%, at least 93%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

[019] In addition to a first capture reagent zone and a second capture reagent zone, a detection zone of a matrix disclosed herein can further comprises one or more additional capture zones. These one or more additional capture zones comprises a competitive ligand to the chemosensory protein comprising the chemosensory protein conjugate of the competitive reagent zone. Like the first capture reagent zone, the one or more additional capture reagent zones serve as an indicator for the presence or absence of the target analyte. In some embodiments, a first capture reagent zone and each of the one or more additional capture reagent zones contain the same amount of the competitive ligand. In some embodiments, a first capture reagent zone and each of the one or more additional capture reagent zones contain a different amount of the competitive ligand. In these later embodiments, the differing amounts can be designed to create a gradient of competitive ligand concentrations. For example, in one embodiment, a capture zone closed to the liquid front of the capillary flow can have a low concentration of competitive ligand and each successive capture zone can have an increased concentration of competitive ligand, such as, e,g,, a doubling amount of increase or a logarithmic increase in the amount. In one embodiment, a capture zone closed to the liquid front of the capillary flow can have a high concentration of competitive ligand and each successive capture zone can have an decreased concentration of competitive ligand, such as, e,g,, a halving amount of decrease or a logarithmic decrease in amount.

[020] A sample application zone of a matrix disclosed herein comprises an area where a sample is initially provided to a lateral device disclosed herein. A sample application zone facilitates uniform application of a sample to a lateral flow device disclosed herein ensuring accurate and consistent results as well as providing standardization when a sample is assayed multiple times using multiple lateral flow devices disclosed herein. [021] An absorbent zone of a matrix disclosed herein comprising an area that facilitates or enhances the flow of a liquid sample across the matrix of a disclosed lateral flow device. An absorbent zone serves as a wick that ensures adequate and uniform capillary flow of a liquid in a timely manner.

[022] In one embodiment, a matrix disclosed herein comprising a sample application zone, a competitive reagent zone, a detection zone, an absorbent zone, or any combination thereof can be composed of the same material. In aspects of this embodiment, a matrix disclosed herein is composed of a nitrocellulose, a nylon, a polyethersulfone, a polyethylenemylar, or a plastic-cast membrane.

[023] A matrix material used for a lateral flow device disclosed herein generally has a capillary rise of between 75 sec/4 cm to 240 sec/4 cm. In aspects of this embodiment, a matrix material used for a lateral flow device disclosed herein has a capillary rise of between 75 sec/4 cm to 240 sec/4 cm, between 80 sec/4 cm to 235 sec/4 cm, between 85 sec/4 cm to 230 sec/4 cm, between 90 sec/4 cm to 225 sec/4 cm, between 100 sec/4 cm to 215 sec/4 cm, between 105 sec/4 cm to 210 sec/4 cm, between 110 sec/4 cm to 205 sec/4 cm, between 115 sec/4 cm to 200 sec/4 cm, between 120 sec/4 cm to 195 sec/4 cm, between 125 sec/4 cm to 190 sec/4 cm, between 130 sec/4 cm to 185 sec/4 cm, between 135 sec/4 cm to 180 sec/4 cm, between 140 sec/4 cm to 175 sec/4 cm, between 145 sec/4 cm to 170 sec/4 cm, between 150 sec/4 cm to 165 sec/4 cm, or between 155 sec/4 cm to 160 sec/4 cm, for example.

[024] In aspects of this embodiment, a matrix material used for a lateral flow device disclosed herein has a capillary rise of between about 75 sec/4 cm to about 240 sec/4 cm, about 80 sec/4 cm to 2 about 35 sec/4 cm, about 85 sec/4 cm to about 230 sec/4 cm, about 90 sec/4 cm to about 225 sec/4 cm, about 100 sec/4 cm to about 215 sec/4 cm, about 105 sec/4 cm to about 210 sec/4 cm, about 110 sec/4 cm to about 205 sec/4 cm, about 115 sec/4 cm to about 200 sec/4 cm, about 120 sec/4 cm to about 195 sec/4 cm, about 125 sec/4 cm to about 190 sec/4 cm, about 130 sec/4 cm to about 185 sec/4 cm, about 135 sec/4 cm to about 180 sec/4 cm, about 140 sec/4 cm to about 175 sec/4 cm, about 145 sec/4 cm to about 170 sec/4 cm, about 150 sec/4 cm to about 165 sec/4 cm, or about 155 sec/4 cm to about 160 sec/4 cm, for example.

[025] In some embodiments, a matrix disclosed herein comprising a sample application zone, a competitive reagent zone, a detection zone, an absorbent zone, or any combination thereof can be composed of different materials. In some embodiments, a sample application zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon. In some embodiments, a competitive reagent zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon. In some embodiments, a detection zone is composed of a nitrocellulose, a nylon, a polyethersulfone, a polyethylenemylar, or a plastic-cast membrane.

[026] A competitive reagent zone and a detection zone disclosed herein generally have a capillary rise of between 75 sec/4 cm to 240 sec/4 cm. In aspects of this embodiment, a competitive reagent zone and a detection zone disclosed herein can have a capillary rise of up to 75 sec/4 cm, up to 80 sec/4 cm, up to 85 sec/4 cm, up to 90 sec/4 cm, up to 100 sec/4 cm, up to 105 sec/4 cm, up to 110 sec/4 cm, up to 115 sec/4 cm, up to 120 sec/4 cm, up to 125 sec/4 cm, up to 130 sec/4 cm, up to 135 sec/4 cm, up to 140 sec/4 cm, up to 145 sec/4 cm, up to 150 sec/4 cm, up to 155 sec/4 cm, up to 160 sec/4 cm, up to 165 sec/4 cm, 170 sec/4 cm, up to 175 sec/4 cm, up to 180 sec/4 cm, up to 185 sec/4 cm, up to 190 sec/4 cm, up to 195 sec/4 cm, up to 200 sec/4 cm, up to 205 sec/4 cm, up to 210 sec/4 cm, up to 215 sec/4 cm, up to 220 sec/4 cm, up to 225 sec/4 cm, up to 230 sec/4 cm, up to 235 sec/4 cm or up to 240 sec/4 cm, for example.

[027] In aspects of this embodiment, a competitive reagent zone and a detection zone disclosed herein can have a capillary rise of at least 75 sec/4 cm, at least 80 sec/4 cm, at least 85 sec/4 cm, at least 90 sec/4 cm, at least 100 sec/4, at least 105 sec/4 cm, at least 110 sec/4 cm, at least 115 sec/4 cm, at least 120 sec/4 cm, at least 125 sec/4 cm, at least 130 sec/4 cm, at least 135 sec/4 cm, at least 140 sec/4 cm, at least 145 sec/4 cm, at least 150 sec/4 cm, at least 155 sec/4 cm or at least 160 sec/4 cm, for example.

[028] A lateral flow device disclosed herein can further include a housing that enclosed the matrix to create a testing device. A testing device can comprise a sample port that allows access to a sample application zone of a matrix where a user can apply a sample to the sample application zone. A testing device can comprise viewing port that allows visual access to a detection zone of a matrix where a user can observe the results of an assay.

[029] The present specification also discloses a testing kit. A testing kit comprises a lateral flow device or a testing device disclosed herein. A testing kit can also include a sample container including a chemosensory protein conjugate disclosed herein. A testing kit can also include instructions on how to use a a lateral flow device or a testing device disclosed herein. [030] In still another aspect, the lateral flow device disclosed herein is provided having the material composition of the sample application zone, the competitive reagent zone, the absorbent zone, or any combination thereof is different from the material composition of the detection zone. In some embodiments, the material composition of the sample application zone can be cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon. In another aspect, the lateral flow devices herein disclosed can have at least one competitive reagent zone comprising cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon. In particular examples, the material composition of the absorbent zone can be cotton fibers or cellulose fibers. In still yet another aspect, the material composition of the detection zone in particular embodiments can be nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm, for example.

[031] In certain aspects, the lateral flow device for detecting the presence or absence of at least one target analyte in a sample can have a solid support that is a laminate backing material of mylar or polyester.

[032] In further aspects, a testing device for determining the presence or absence of a target analyte in a liquid sample is herein disclosed, where the testing device comprises a lateral flow device as described herein and a housing that encloses the matrix. In particular configurations the housing includes a sample port and viewing port.

[033] In yet another aspect, a testing kit for determining the presence or absence of a target analyte in a liquid sample is herein disclosed, the testing kit comprising a lateral flow device as herein disclosed; or a testing device as disclosed above having a housing enclosing the matrix and in one embodiment, the housing further comprising a sample and a viewing port; the testing kit further comprising a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

[034] The kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample. In particular embodiments, the the chemosensory protein conjugate of the testing kit is lyophilized or freeze-dried. In further embodiments of the testing kits described herein and provided in accordance with the teachings of the present invention, the sample container further comprises a culture media capable of supporting bacterial growth.

[035] In still another aspect, a testing container for determining the presence or absence of a target analyte in a liquid sample is disclosed herein, the testing container comprising a first compartment comprising a lateral flow device as herein disclosed (e.g. those lateral flow devices exemplarily described above), or a testing device as herein disclosed (e.g. those testing devices exemplarily described above) and a second compartment comprising a sample region and, further, a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition. The seal covering the channel is provided as intact, the seal preventing fluid communication between the first and second compartments via the channel, whereas when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur/is established. In particular embodiments, the sample region of the second compartment includes a culture media capable of supporting bacterial growth.

[036] Further aspects of the present specification disclose methods of determining the presence or absence of a target analyte in a liquid sample utilizing the lateral flow devices herein disclosed, a testing device as herein disclosed or a testing container as herein disclosed, the method comprising the steps of contacting a liquid sample to a lateral flow device, a testing device, a testing kit or a testing container as herein disclosed, permitting the liquid sample to flow through the chemosensory protein conjugate zone and the detection reagent zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone. Observation of the detection zone is conducted to determine to determine the presence or absence of analyte in the liquid sample. Visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample and visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample. Is some embodiments, the methods disclose herein determine the presence or absence of a coliform bacteria and/or fecal contamination. In aspects of this embodiment, the disclosed methods detect the presence of indole or a derivative of indole as a marker for a coliform bacteria and/or fecal contamination. [037] Exemplary method steps are disclosed herein for determining the presence or absence of a target analyte in a liquid sample. In one aspect, a method includes the steps of mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof, contacting the liquid sample to a lateral flow device, a testing device, a testing kit, or a testing container as herein disclosed. The liquid sample is permitted to flow through the detection reagent zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone. Observation of the detection zone is conducted to determine to determine the presence or absence of analyte in the liquid sample. Visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample and visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

[038] Further exemplary method steps are disclosed herein for determining the presence or absence of a target analyte in a liquid sample. In another aspect, a method includes the steps of adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample- media mixture and incubating the sample-media mixture for a period of time. The sample-media mixture is then contacted to a lateral flow device, a testing device, a testing kit, or a testing container as herein disclosed. The sample-media mixture is permitted to flow through the detection reagent zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone. Observation of the detection zone is conducted to determine to determine the presence or absence of analyte in the liquid sample. Visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample and visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

[039] In one aspect, a method for determining the presence or absence of a target analyte in a liquid sample is herein disclosed, the method comprising the steps of adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture, incubating the sample-media mixture for a period of time and adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof. This is followed by contacting the conjugate mixture to a lateral flow device, a testing device, a testing kit, or a testing container as herein disclosed and permitting the conjugate mixture to flow through the detection reagent zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone. Observation of the detection zone is conducted to determine to determine the presence or absence of analyte in the liquid sample. Visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample and visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

[040] In particular embodiments, the methods include the step of incubating sample-media mixture for a time that is at most 1 hours, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours. In particular embodiments incubation times can be at least about 15 minutes, least about 30 minutes, least about 60 minutes, least about 90 minutes least about 120 minutes, least about 180 minutes, least about 240, minutes least about 300 minutes, least about 330 minutes or at least about 390 minutes.

[041] It is to be understood that in any one of the embodiments of the lateral flow device, a testing device, a testing kit, or a testing container or method of use related thereto and as herein disclosed, the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte can be detected. Further, it is to be understood that the liquid sample utilized with any lateral flow device, testing device, testing kit, testing container or method of use related thereto and as herein disclosed, can be a water sample.

[042] Aspects of the present specification can be described as follows:

1. A lateral flow device for detecting the presence or absence of a target analyte in a sample comprising: a) a matrix that supports the flow of a liquid sample comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate.

The lateral flow device of embodiment 1 , wherein the plurality of capture reagent zones further comprise one or more additional capture reagent zones, each of the one or more additional capture zones comprising a competitive ligand, the one or more additional capture zones each serving as an indicator for the presence or absence of the target analyte to the chemosensory protein.

The lateral flow device of any one of embodiment 1 or 2, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain the same amount of the competitive ligand.

The lateral flow device of embodiments 1 -3, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain a different amount of the competitive ligand.

The lateral flow device of embodiments 1-4, wherein the different amount of the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones established a concentration gradient.

The lateral flow device of any one of embodiments 1-5, wherein the matrix further comprises a sample application zone, an absorbent zone, or both a sample application zone and an absorbent zone.

The lateral flow device of any one of embodiments 1-6, wherein the chemosensory protein is an odorant binding protein.

The lateral flow device of embodiment 7, wherein the odorant binding protein is an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33.

The lateral flow device of embodiment 7 or 8, wherein the odorant binding protein is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

The lateral flow device of any one of embodiment 1-9, wherein nanoparticle is a colloidal gold particle, a colored latex particle, a carbon particle, a selenium particle, a chemiluminescent particle, a bioluminescent particle, a fluorescent particle, a quantum dot particle, an upconverting phosphor particle, a liposome including a dye.

The lateral flow device of any one of embodiments 1-10, wherein nanoparticle is about 15 nm to about 800 nm in size.

The lateral flow device of any one of embodiments 1-11, wherein the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones is conjugated to a carrier protein.

The lateral flow device of embodiment 12, wherein the carrier protein is a serum albumin or a thyroglobin.

The lateral flow device of any one of embodiments 1-13, wherein the competitive ligand is indole, 3-methyl indole or another derivative of indole.

The lateral flow device of any one of embodiments 1-14, wherein the sample application zone, the competitive reagent zone, the detection zone, and the absorbent zone are each composed of a material that is the same.

The lateral flow device of embodiment 15, wherein the material is nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

The lateral flow device of any one of embodiments 1-14, wherein the sample application zone, the competitive reagent zone, the absorbent zone, or any combination thereof is composed of a material that is different from the material comprising the detection zone.

The lateral flow device of embodiment 17, wherein the sample application zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon.

The lateral flow device of embodiment 17, wherein the competitive reagent zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon.

The lateral flow device of embodiment 17, wherein the absorbent zone is composed of cotton fibers or cellulose fibers.

The lateral flow device of embodiment 17, wherein the detection zone is composed of nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

The lateral flow device of any one of embodiments 1-21, wherein the device further comprises a solid support.

The lateral flow device of embodiment 22, wherein the solid support is a laminate backing material mylar or polyester. A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device as defined in embodiments 1-23; b) a housing that encloses the matrix.

The testing device of embodiment 24, wherein the housing further comprises a sample port and viewing port.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-23 or a testing device of embodiment 24 or 25; and b) a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-23 or a testing device of embodiment 24 or 25; and b) a sample container.

The testing kit of embodiment 26 or 27, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample.

The testing kit of any one of embodiments 26-28, wherein the chemosensory protein conjugate is lyophilized or freeze-dried.

The testing kit of any one of embodiments 26-29, wherein the sample container further comprises a culture media capable of supporting bacterial growth.

A testing container for determining the presence or absence of a target analyte in a liquid sample, the testing container comprising: a) a first compartment comprising a lateral flow device as defined in embodiments 1-23 or a testing device of embodiment 24 or 25; b) a second compartment comprising a sample region; and c) a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition; wherein when the seal covering the channel is intact, the seal prevents fluid communication between the first and second compartments via the channel; and wherein when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur.

The testing container of embodiment 31, wherein the sample region includes a culture media capable of supporting bacterial growth.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) contacting a liquid sample to a lateral flow device as defined in any one of embodiments 1-23, a testing device according to embodiment 24 or 25, a testing kit as defined in any one of embodiments 26-30, or a testing container as defined in embodiments 31 or 32; b) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and c) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; b) contacting the liquid sample to a lateral flow device as defined in any one of embodiments 1-23, a testing device according to embodiment 24 or 25, a testing kit as defined in any one of embodiments 26-30, or a testing container as defined in embodiments 31 or 32; c) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and d) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) contacting the sample-media mixture to a lateral flow device as defined in any one of embodiments 1-23, a testing device according to embodiment 24 or 25, a testing kit as defined in any one of embodiments 26-30, or a testing container as defined in embodiments 31 or 32; d) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and e) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; d) contacting the conjugate mixture to a lateral flow device as defined in any one of embodiments 1 -23, a testing device according to embodiment 24 or 25, a testing kit as defined in any one of embodiments 26-30, or a testing container as defined in embodiments 31 or 32; e) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and f) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

The method of embodiment 35 or 36, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours.

The method of any one of embodiments 32-36, wherein the method can detect the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte.

The method of any one of embodiments 32-37, wherein the liquid sample is a water sample. [043] Aspects of the present specification can be described as follows:

1. A lateral flow device for detecting the presence or absence of a target analyte in a sample comprising: a) a matrix that supports the flow of a liquid sample comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a solid support.

2. The lateral flow device of embodiment 1 , wherein the plurality of capture reagent zones further comprise one or more additional capture reagent zones, each of the one or more additional capture zones comprising a competitive ligand, the one or more additional capture zones each serving as an indicator for the presence or absence of the target analyte to the chemosensory protein.

3. The lateral flow device of any one of embodiment 1 or 2, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain the same amount of the competitive ligand.

4. The lateral flow device of embodiments 1 -3, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain a different amount of the competitive ligand.

5. The lateral flow device of embodiments 1-4, wherein the different amount of the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones established a concentration gradient.

6. The lateral flow device of any one of embodiments 1-5, wherein the matrix further comprises a sample application zone, an absorbent zone, or both a sample application zone and an absorbent zone.

7. The lateral flow device of any one of embodiments 1-6, wherein the chemosensory protein is an odorant binding protein.

8. The lateral flow device of embodiment 7, wherein the odorant binding protein is an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33. The lateral flow device of embodiment 7 or 8, wherein the odorant binding protein is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

The lateral flow device of embodiment 17, wherein the absorbent zone is composed of cotton fibers or cellulose fibers. The lateral flow device of embodiment 17, wherein the detection zone is composed of nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

The lateral flow device of any one of embodiments 1-21, wherein the solid support is a laminate backing material mylar or polyester.

A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device as defined in embodiments 1-22; b) a housing that encloses the matrix.

The testing device of embodiment 23, wherein the housing further comprises a sample port and viewing port.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-22 or a testing device of embodiment 23 or 24; and b) a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-22 or a testing device of embodiment 23 or 34; and b) a sample container.

The testing kit of embodiment 25 or 26, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample.

The testing kit of any one of embodiments 25-27, wherein the chemosensory protein conjugate is lyophilized or freeze-dried.

The testing kit of any one of embodiments 25-28, wherein the sample container further comprises a culture media capable of supporting bacterial growth.

A testing container for determining the presence or absence of a target analyte in a liquid sample, the testing container comprising: a) a first compartment comprising a lateral flow device as defined in embodiments 1 -22 or a testing device of embodiment 23 or 24; b) a second compartment comprising a sample region; and c) a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition; wherein when the seal covering the channel is intact, the seal prevents fluid communication between the first and second compartments via the channel; and wherein when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur. The testing container of embodiment 30, wherein the sample region includes a culture media capable of supporting bacterial growth.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) contacting a liquid sample to a lateral flow device as defined in any one of embodiments 1-22, a testing device according to embodiment 23 or 24, a testing kit as defined in any one of embodiments 25-29, or a testing container as defined in embodiments 30 or 31 ; b) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and c) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; b) contacting the liquid sample to a lateral flow device as defined in any one of embodiments 1-22, a testing device according to embodiment 23 or 24, a testing kit as defined in any one of embodiments 25-29, or a testing container as defined in embodiments 30 or 31; c) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and d) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) contacting the sample-media mixture to a lateral flow device as defined in any one of embodiments 1-22, a testing device according to embodiment 23 or 24, a testing kit as defined in any one of embodiments 25-29, or a testing container as defined in embodiments 30 or 31; d) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and e) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; d) contacting the conjugate mixture to a lateral flow device as defined in any one of embodiments 1-22, a testing device according to embodiment 23 or 24, a testing kit as defined in any one of embodiments 25-29, or a testing container as defined in embodiments 30 or 31; e) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and f) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

The method of embodiment 33 or 34, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours. 36. The method of any one of embodiments 31-35, wherein the method can detect the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte.

37. The method of any one of embodiments 31-36, wherein the liquid sample is a water sample.

[044] Aspects of the present specification can be described as follows:

1. A lateral flow device for detecting the presence or absence of a target analyte in a sample, the device comprising: a) a matrix that supports the flow of a liquid sample comprising i) a sample application zone; ii) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and iii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; iv) an absorbent zone; and b) a solid support.

6. The lateral flow device of any one of embodiments 1-5, wherein the chemosensory protein is an odorant binding protein. The lateral flow device of embodiment 6, wherein the odorant binding protein is an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33.

The lateral flow device of embodiment 6 or 7, wherein the odorant binding protein is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

The lateral flow device of any one of embodiments 1-8, wherein nanoparticle is a colloidal gold particle, a colored latex particle, a carbon particle, a selenium particle, a chemiluminescent particle, a bioluminescent particle, a fluorescent particle, a quantum dot particle, an upconverting phosphor particle, a liposome including a dye.

The lateral flow device of any one of embodiments 1-9, wherein nanoparticle is about 15 nm to about 800 nm in size.

The lateral flow device of any one of embodiments 1-10, wherein the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones is conjugated to a carrier protein.

The lateral flow device of embodiment 11, wherein the carrier protein is a serum albumin or a thyroglobin.

The lateral flow device of any one of embodiments 1-12, wherein the competitive ligand is indole, 3-methyl indole or another derivative of indole.

The lateral flow device of any one of embodiments 1-13, wherein the sample application zone, the competitive reagent zone, the detection zone, and the absorbent zone are each composed of a material that is the same.

The lateral flow device of embodiment 14, wherein the material is nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

The lateral flow device of any one of embodiments 1-13, wherein the sample application zone, the competitive reagent zone, the absorbent zone, or any combination thereof is composed of a material that is different from the material comprising the detection zone.

The lateral flow device of embodiment 16, wherein the sample application zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon.

The lateral flow device of embodiment 16, wherein the competitive reagent zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon. The lateral flow device of embodiment 16, wherein the absorbent zone is composed of cotton fibers or cellulose fibers.

The lateral flow device of embodiment 16, wherein the detection zone is composed of nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

The lateral flow device of any one of embodiments 1 -20, wherein the solid support is a laminate backing material mylar or polyester.

A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device as defined in embodiments 1-21; b) a housing that encloses the matrix.

The testing device of embodiment 22, wherein the housing further comprises a sample port and viewing port.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-21 or a testing device of embodiment 22 or 23; and b) a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in embodiments 1-21 or a testing device of embodiment 22 or 23; and b) a sample container.

The testing kit of embodiment 24 or 25, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample.

The testing kit of any one of embodiments 24-26, wherein the chemosensory protein conjugate is lyophilized or freeze-dried.

The testing kit of any one of embodiments 24-27, wherein the sample container further comprises a culture media capable of supporting bacterial growth.

A testing container for determining the presence or absence of a target analyte in a liquid sample, the testing container comprising: a) a first compartment comprising a lateral flow device as defined in embodiments 1 -21 or a testing device of embodiment 22 or 23; b) a second compartment comprising a sample region; and c) a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition; wherein when the seal covering the channel is intact, the seal prevents fluid communication between the first and second compartments via the channel; and wherein when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur.

The testing container of embodiment 29, wherein the sample region includes a culture media capable of supporting bacterial growth.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) contacting a liquid sample to a lateral flow device as defined in any one of embodiments 1-21, a testing device according to embodiment 22 or 23, a testing kit as defined in any one of embodiments 24-28, or a testing container as defined in embodiments 29 or 30; b) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and c) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; b) contacting the liquid sample to a lateral flow device as defined in any one of embodiments 1-21, a testing device according to embodiment 22 or 23, a testing kit as defined in any one of embodiments 24-28, or a testing container as defined in embodiments 29 or 30; c) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and d) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) contacting the sample-media mixture to a lateral flow device as defined in any one of embodiments 1-21, a testing device according to embodiment 22 or 23, a testing kit as defined in any one of embodiments 24-28, or a testing container as defined in embodiments 29 or 30; d) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and e) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; d) contacting the conjugate mixture to a lateral flow device as defined in any one of embodiments 1 -21 , a testing device according to embodiment 22 or 23, a testing kit as defined in any one of embodiments 24-28, or a testing container as defined in embodiments 29 or 30; e) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and f) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample. 35. The method of embodiment 33 or 34, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours.

36. The method of any one of embodiments 31-35, wherein the method can detect the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte.

[045] Aspects of the present specification can be described as follows:

1. A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a housing that encloses the matrix.

2. The testing device of embodiment 1, wherein the housing further comprises a sample port and viewing port.

[046] Aspects of the present specification can be described as follows:

1. A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a housing that encloses the matrix.

2. The device of embodiment 1 , wherein the housing further comprises a sample port and viewing port. [047] Aspects of the present specification can be described as follows:

1. A testing kit for determining the presence or absence of a target analyte in a liquid sample, the kit comprising: a) a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

2. The kit of embodiment 1, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample.

3. The kit of embodiment 1 or 2, wherein the chemosensory protein conjugate is lyophilized or freeze-dried.

4. The kit of any one of embodiments 1-3, wherein the sample container further comprises a culture media capable of supporting bacterial growth.

[048] Aspects of the present specification can be described as follows:

1. A testing kit for determining the presence or absence of a target analyte in a liquid sample, the kit comprising: a) a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a sample container.

2. The kit of embodiment 1, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample. 3. The kit of embodiment 1 or 2, wherein the sample container includes a culture media capable of supporting bacterial growth.

[049] Aspects of the present specification can be described as follows:

1. A testing container for determining the presence or absence of a target analyte in a liquid sample, the container comprising: a) a first compartment comprising a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; and b) a second compartment comprising a sample region; and c) a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition; wherein when the seal covering the channel is intact, the seal prevents fluid communication between the first and second compartments via the channel; and wherein when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur.

2. The container of embodiment 1, wherein the sample region includes a culture media capable of supporting bacterial growth.

[050] Aspects of the present specification can be described as follows:

1. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) contacting a liquid sample to a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; b) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and c) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

2. The method of embodiment 1, wherein the method can detect the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte.

[051] Aspects of the present specification can be described as follows:

1. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; b) contacting the liquid sample to a lateral flow device comprising a matrix that supports the flow of a liquid sample, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; c) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and d) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

[052] Aspects of the present specification can be described as follows:

1. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) contacting the sample-media mixture to a lateral flow device comprising a matrix that supports the flow of the sample-media mixture, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; d) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and e) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

3. The method of embodiment 1, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours. [053] Aspects of the present specification can be described as follows:

1. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; d) contacting the conjugate mixture to a lateral flow device comprising a matrix that supports the flow of the conjugate mixture, the matrix comprising i) a competitive reagent zone including a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle; and ii) a detection zone including a plurality of capture reagent zones, the plurality of capture reagent zones including a first capture reagent zone comprising a competitive ligand to the chemosensory protein, the first capture reagent zone serving as an indicator for the presence or absence of the target analyte to the chemosensory protein and a second capture reagent zone comprising an antibody having specificity for the chemosensory protein, the second capture reagent zone serving as a positive control for the chemosensory protein conjugate; e) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and f) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

3. The method of embodiment 1 or 2, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours.

EXAMPLES [054] The following non-limiting examples are provided for illustrative purposes only in order to facilitate a more complete understanding of representative embodiments now contemplated. These examples should not be construed to limit any of the embodiments described in the present specification, including those pertaining to the compounds, devices, or methods and uses disclosed herein.

[055] Here embodiments utilizing a particular odorant binding protein, AgamOBPl, from the mosquito was utilized in providing biosensor devices in accordance with the present disclosure that were able to detect a characteristic bacterial metabolite, indole.

[056] Cloning of AgamOBPl. A PCR-amplified DNA fragment encoding AgamOBPl (AF437884) was cloned into pRSET-B (Thermo Fisher Scientific, Waltham, USA) and soluble recombinant protein (rAgamOBPl) was produced in E. coli BL21 Star (DE3)pLysS cells. The rAgamOBPl protein was purified on a nickel-NTA column following the manufacturer’s directions (Thermo Fisher Scientific, Waltham, USA), eluted with 5 mM EDTA and subjected to extensive dialysis against 50 mM Tris-HCl pH 7.4.

[057] An Attenu assay system. The Attenu assay takes advantage of the fluorescent properties of the dye, 1-NPN (N-Phenyl-l-naphthylamine, CAS 90-30-2). 1-NPN exhibits a detectably altered emission spectrum when interacting with the ligand-binding pocket of insect OBPs in that the peak emission wavelength is shifted from 460 nm to 416 nm and the maximum response amplitude is increased. When a ligand displaces 1-NPN from the OBP’s binding pocket, fluorescent response is reduced. This fluorescence quenching can be detected using a spectrophotometer. The Attenu screening system was utilized with concentrations of ligands and rAgamOBPl in the mM range and fluorescence was detected using a Molecular Devices Gemini XPS spectrofluorometer (Sunnyvale, CA, USA).

[058] Lateral flow devices. The lateral flow devices utilized absorbent pads supporting a nitrocellulose membrane that is in contact with the sample and conjugate pads. The conjugate pad contains rAgamOBPl conjugated to 30 nm colloidal gold, which served as a source of color for both test and control lines; colloidal gold is visible to the naked eye. The test line contains a competitive ligand and the control line contains an anti-r AgamOBPl antibody. If a tested sample contains indole or its derivatives, these molecules will displace the competitive ligand from the test line; a positive result is one in which the visible test line is lost. The control line will verify functionality of the device if a visible signal is produced as the anti-rAgamOBPl antibodies capture displaced rAgamOBPl. The device is depicted in Figure 4.

[059] Devices are assembled from sheets that are striped with the appropriate molecules using a SynQuad Automated Dispenser (Cartesian Technologies, Irvine, USA) and cut into 5 mm strips. The strips can be supported by an inert, rigid material such as plastic (e.g., polypropylene).

[060] A tested solution travels along the strip via capillary action with the speed of flow determined by the pore size of the membrane. Visible results appear in less than 20 minutes. More detailed analyses of the results can be achieved if the strips are allowed to dry for 24 hours and then read using a Qiagen ESE-Quant GOLD scanner (Qiagen, Hilden, Germany) to determine the absorbance by position in 40 pm increments. The scanner merely allows for quantification as well as detection of differences not obvious to the naked eye; it is not necessary for operation of the device or rapid detection of indole.

[061] In order to generate a test strip capable of acting as a real-time positive control, we used the Attenu assay to isolate a synthetic ligand for rAgamOBPl from a combinatorial chemical library (ChemBridge, San Diego, USA). The synthetic ligand was used in the positive control strip. Tests were performed using a 2X ligand dilution series in which strips are exposed to differing concentrations of indolepropionic acid bound to BSA (IPA-BSA) with constant amounts of Hi- Veg media (containing high levels of tryptophan, the metabolic breakdown of which produces indole) and of AgamOBPl conjugated to 30 nm colloidal gold.

[062] The completed device was evaluated using E. coli K-12 cells (ATCC, Manassas, USA). Liquid cultures of K-12 were diluted 1000X in Hi-Veg Media to provide a source of tryptophan that the cells metabolize into indole. Cells were incubated for 60 minutes and 500 pi aliquots were removed every 30 minutes for testing. Tests lasted 3 hours and the absorbance at OD600 of tested samples from ranged 0.005 A to 0.05 A.

[063] Fluorescence quenching assay to detect coliform bacteria and fecal contamination. The Attenu assay system was used as the platform for a plate-based biosensor with recombinant AgamOBPl at 1 pM as the detector element. This device was tested with samples of bacterial cells at various concentrations as well as aqueous solutions containing indole. [064] Indole detection testing was performed with varying concentrations of indole in water. Initial results indicated that the biosensor detected indole at concentrations less than 100 nM (~5 ppb; Figure 5) and detection requires 30 minutes or less, making this biosensor not only more sensitive than the standard method of detection but also much more rapid. Expanded testing using indole derivatives including 1 -methyl indole, 3 -methyl indole (skatole), and 5 -methyl indole each at 10 pM concentrations showed binding to skatole only (Figure 1).

[065] Figure 1. A novel plate-based biosensor to detect indole with high sensitivity. The Attenu fluorescence-quenching assay was adapted to develop a biosensor and rAgamOBPl at 1 mM served as the detector element. The fluorescent dye, 1-NPN, binds rAgamOBPl and results in a signal that can be detected spectrophotometrically. Analytes capable of binding to rAgamOBPl displace the dye from the protein’s binding pocket, thus resulting in fluorescence quenching. The tested compounds at 10 mM concentration included indole (Ind), 3-methyl indole (3-Me Ind), 1 -methyl indole (1-Me Ind), and 5-methyl indole (5-Me Ind). Six replicas of each compound were evaluated. Three replicates were performed with rAgamOBPl and buffer alone as controls. The loss of signal indicates a binding event; thus, note the signal drop generated by indole and to a lesser degree 3- methyl indole. The bars are STDEV determined with Excel and the raw data are shown in Table 1.

These are raw numbers and statistical analysis of data shown in Fig. 1. Area under the response curve fromt eh Attenu assay is shown in the first six rows. The indole derivative is listed on top. Averages and statistical analysis are shown in the last 3 rows.

[066] Detecting E. coli cells. E. coli detection was tested for response time and sensitivity. The plate-based biosensor detected 50 cfu (colony forming units) of E. coli strain K-12 cells after a 30- minute incubation, and the signal became increasingly robust up to the maximum incubation time of 120 minutes (Figure 2). The plate-based biosensor was also tested for sensitivity, and detected less than 5 colony -forming units (cfu) of E. cob after a 30-minute incubation (Figure 2).

[067] Figure 2. Rapidly detecting E. cob cells. The biosensor based on the Attenu fluorescence quenching assay utilized rAgamOBPl at 1 mM as the detector element. Data were collected using a spectrophotometer. Bar graph of the areas under the response curves for the control (media, no bacteria); 5 cfu of K-12 (green); 50 cfu of K-12 (red); and 500 cfu of an E. coli strain (MG1651, Thermo Fisher Scientific, Waltham, USA) without functional tryptophanase and therefore unable to produce indole (purple). The black bars are STDEV plotted with Excel and the raw data are shown in Table 2.

Table 2

Raw numbers and statistical analysis of the data shown in Fig. 2. Area under the response curve from the Attenu assay is shown in the first 3 rows. The type of bacteria is listed above the table. Averages and statistical analysis is shown in the last 3 row.

[068] Detecting fecal contamination. The plate-based biosensor was also tested for the ability to detect fecal contamination in an aqueous solution. Canine feces were diluted in water at abundances ranging from 100 ppm (parts per million) to 10 ppb (parts per billion). The biosensor was effective for the detection of fecal contamination in water with a reliable detection limit of 100 ppb. The linear range of detection is from 10 ppm and 100 ppb (Figure 3), making this biosensor a responsive, sensitive instrument for fecal contamination of water supplies.

[069] Figure 3. Detecting fecal contamination of water. Data were obtained using the plate-based biosensor with rAgamOBPl at 1 mM as the detector element. Data were collected using a spectrophotometer. Control samples contain only water. The areas under each curve plotted on a semi-logarithmic scale in order to evaluate the linearity of the biosensor’s response. The linear range of detection ranges from 10 ppm to 100 ppb. The red bars are STDEV plotted with Excel and the raw data are shown in Table 3.

Table 3

Raw numbers and statistical analysis of the data shown in Fig. 3. Area under the response curve from replicates in the Attenu assay is shown in the first three rows. The concentration of feces tested is listed at the top. Averages and statistical analysis are shown in the last 3 rows.

[070] Low-cost portable lateral flow device to detect fecal contamination. Lateral flow devices were assembled in a format resembling commercially available pregnancy test kits with blank “cassettes” made of inert plastic (e.g., polypropylene). The device is based on competitive binding between an analyte in the tested sample and a synthetic ligand to rAgamOBPl and results are interpreted based on the color of a test line and a control line. The device detected indole in aqueous solutions at concentrations as low as 100 ppm (Figure 4).

[071] Figure 4. Lateral flow biosensor to detect indole. The detection scheme utilizes rAgamOBPl that is rendered visible to the naked eye when it is conjugated colloidal gold. Thus, rAgamOBPl -colloidal gold conjugate lines were striped onto a nitrocellulose membrane sandwiched between sample and conjugate pads. Detection of indole is based on the competition between any indole present in a given sample to be tested and the synthetic ligand in the test line. A positive result is reflected by the loss of a visible test line on the device and indicates that indole is present in the tested sample. Therefore, the device displays only the control line for a positive result. When testing a sample lacking indole the device displays two lines - the test line and the control line (a) Cut-away diagram of a standard lateral flow device (b) The indole biosensor device. The sample that was added to the upper cassette (-) contained only PBS and shows how a negative result appears (2 lines visible). The sample added to the lower cassette (+) contained PBS plus indole (lOOppm) and demonstrates a positive result (only control line visible) (c) & (d) Operating principles of the lateral flow indole biosensor. In (c) the control (PBS) result is shown. Since no ligand is present in the analyte the rAgamOBPl-colloidal gold conjugate is free to bind the test line as well as the control line. A positive result is shown in (d). The indole in the analyte binds the rAgamOBPl-colloidal gold conjugate and prevents binding to the test line; therefore, only the control line is visible. [072] Figure 5. Response of AgamOBPl to various dilutions of indole using the Attenu assay. Concentrations of indole including, from left to right, 0, 100 nM, 1 mM, 2.5 mM, 5 pM, and 10 pM were tested with the Attenu assay. Shown on the plot are two separate experiments run with the same indole dilutions. The first experiment is shown as blue circles, the second as red diamonds. Note that a drop in response is visible even at 100 nM. Response of AgamOBPl to various dilutions of indole using the Attenu assay of indole including, from left to right, 0, 100 nM, 1 pM, 2.5 pM, 5 pM, and 10 pM were tested with the Attenu assay. Shown on the plot are two separate experiments run with the same indole dilutions. The first experiment is shown as blue circles, the second as red diamonds drop in response is visible even at 100 nM.

[073] Herein we describe a novel sensor technology encompassing an olfactory protein from Anopheles gambiae mosquitoes, AgamOBPl, as the detector element. AgamOBPl binds analytes associated with coliform bacteria and does so with high specificity and sensitivity, allowing the rapid detection of low level E. coli contamination in water supplies. Moreover, AgamOBPl is a resilient protein that has the typically robust OBP structure with six a-helices that are stabilized by disulfide bridges; in our laboratory, samples of AgamOBPl remained active after being stored in 50 mM Tris-HCl pH 7.4 for up to 5 years. We demonstrate two implementations of AgamOBPl as a detector element in biosensors that indicate the presence of coliform bacterial metabolites in aqueous solutions.

[074] One implementation of an OBP-based biosensor is based on an established reporter mechanism, the Attenu assay, and can be implemented in vitro (Figures 1-3). In this implementation, based on competitive binding between either a fluorescent indicator dye or any given analyte for AgamOBPl’ s binding pocket, recombinantly expressed AgamOBPl can detect the bacterial metabolite, indole, in concentrations below 1 mM in aqueous solutions (Appendix, Figure Al). The plate-based biosensor can also detect E. coli cells rapidly, requiring less than 30 minutes for a result (Figure 2).

[075] However, insect OBP-based biosensors can also be incorporated into lateral flow devices assembled as nitrocellulose strips or sheets with a paper sample pad or wick and supported on a plastic (e.g., PVC) substrate. Such devices are similar in implementation to commonly available pregnancy tests or“dipsticks”, and can take advantage of existing packaging facilities for mass production purposes as a result. In this biosensor a test strip contains a known ligand for AgamOBPl, and a control strip contains an antibody against AgamOBPl. When an aqueous analyte mixture is introduced, an AgamOBPl -colloidal gold complex travels laterally along the device’s surface. If AgamOBPl has bound none of the ligands from the tested aqueous solution, then it will bind the test strip, deposit colloidal gold, and cause a color change. A ligand from the tested solution bound by AgamOBPl will prevent this color change in the test strip; thus, the absence of a color change on the test strip indicates a positive result. The control strip verifies that AgamOBPl is present and travelled laterally along the device.

[076] Both implementations of insect OBP-based biosensors yield data in less than 1 hour. Our results indicate the OBP-based approach is at least one thousand times (1000X) more sensitive than previous methods (e.g., the indole spot test) used for determining the presence of indole. The devices presented herein thus provide proof of concept for insect OBP-based biosensors in general. Future OBP-based biosensors can take advantage of the high sensitivity and versatility of the insect chemosensory system to detect a wide variety of analytes.

[077] The techniques described here can be used to assemble OBP-based biosensors for a wide variety of applications including the detection of environmental, chemical, or biological compounds or contaminants. Such uses include the detection of toxins or stereoisomers generated during chemical or pharmaceutical synthesis, the detection of harmful volatile organic compounds (VOCs), quality control of foods and pharmaceuticals, and the detection of volatile compounds present in weapons or explosives. These biosensors can also be used in medical diagnostics as well as numerous other applications where high detector stability, high sensitivity and analyte selectivity are required. When used as detectors of coliform bacteria in aqueous solutions, the OBP-based biosensors described have specific advantages over bacterial culture- or plate-based detection methods in that the latter can only reveal the presence of living cells and require up to 24 hours to do so. OBP-based detectors are not only capable of rapid detection but can also be targeted against coliform-specific metabolic byproducts - that is, the biosensors detect coliform contamination whether the sample contains living cells or not. Unlike PCR-based methods, OBP- based biosensors can be implemented as simple devices that do not require high levels of end-user expertise. Furthermore, although antibody-based biosensors are established they are limited to detecting analytes that are sufficiently antigenic. Since OBP-based detector elements do not rely on antibodies, they can detect analytes with poor antigenic properties. Thus, the described platform technology has immediate application to a variety of important sensor and detector implementations. [078] In closing, it is to be understood that although aspects of the present specification are highlighted by referring to specific embodiments, one skilled in the art will readily appreciate that these disclosed embodiments are only illustrative of the principles of the subject maher disclosed herein. Therefore, it should be understood that the disclosed subject maher is in no way limited to a particular compound, composition, article, apparatus, methodology, protocol, and/or reagent, etc., described herein, unless expressly stated as such. In addition, those of ordinary skill in the art will recognize that certain changes, modifications, permutations, alterations, additions, subtractions and sub-combinations thereof can be made in accordance with the teachings herein without departing from the spirit of the present specification. It is therefore intended that the following appended claims and claims hereafter introduced are interpreted to include all such changes, modifications, permutations, alterations, additions, subtractions and sub-combinations as are within their true spirit and scope.

[079] Certain embodiments of the present invention are described herein, including the best mode known to the inventors for carrying out the invention. Of course, variations on these described embodiments will become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventor expects skilled artisans to employ such variations as appropriate, and the inventors intend for the present invention to be practiced otherwise than specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject maher recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described embodiments in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

[080] Groupings of alternative embodiments, elements, or steps of the present invention are not to be construed as limitations. Each group member may be referred to and claimed individually or in any combination with other group members disclosed herein. It is anticipated that one or more members of a group may be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[081] Unless otherwise indicated, all numbers expressing a characteristic, item, quantity, parameter, property, term, and so forth used in the present specification and claims are to be understood as being modified in all instances by the term“about.” As used herein, the term “about” means that the characteristic, item, quantity, parameter, property, or term so qualified encompasses a range of plus or minus ten percent above and below the value of the stated characteristic, item, quantity, parameter, property, or term. Accordingly, unless indicated to the contrary, the numerical parameters set forth in the specification and attached claims are approximations that may vary. For instance, as mass spectrometry instruments can vary slightly in determining the mass of a given analyte, the term "about" in the context of the mass of an ion or the mass/charge ratio of an ion refers to +/-0.50 atomic mass unit. At the very least, and not as an attempt to limit the application of the doctrine of equivalents to the scope of the claims, each numerical indication should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[082] Use of the terms“may” or“can” in reference to an embodiment or aspect of an embodiment also carries with it the alternative meaning of“may not“ or“cannot.” As such, if the present specification discloses that an embodiment or an aspect of an embodiment may be or can be included as part of the inventive subject matter, then the negative limitation or exclusionary proviso is also explicitly meant, meaning that an embodiment or an aspect of an embodiment may not be or cannot be included as part of the inventive subject matter. In a similar manner, use of the term“optionally” in reference to an embodiment or aspect of an embodiment means that such embodiment or aspect of the embodiment may be included as part of the inventive subject matter or may not be included as part of the inventive subject matter. Whether such a negative limitation or exclusionary proviso applies will be based on whether the negative limitation or exclusionary proviso is recited in the claimed subject matter.

[083] Notwithstanding that the numerical ranges and values setting forth the broad scope of the invention are approximations, the numerical ranges and values set forth in the specific examples are reported as precisely as possible. Any numerical range or value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements. Recitation of numerical ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate numerical value falling within the range. Unless otherwise indicated herein, each individual value of a numerical range is incorporated into the present specification as if it were individually recited herein. [084] The terms“a,”“an,”“the” and similar references used in the context of describing the present invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. Further, ordinal indicators - such as“first,”“second,”“third,” etc. - for identified elements are used to distinguish between the elements, and do not indicate or imply a required or limited number of such elements, and do not indicate a particular position or order of such elements unless otherwise specifically stated. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g.,“such as”) provided herein is intended merely to better illuminate the present invention and does not pose a limitation on the scope of the invention otherwise claimed. No language in the present specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[085] When used in the claims, whether as filed or added per amendment, the open-ended transitional term“comprising”, variations thereof such as “comprise” and“comprises”, and equivalent open-ended transitional phrases thereof like“including,”“containing” and“having”, encompasses all the expressly recited elements, limitations, steps, integers, and/or features alone or in combination with unrecited subject matter; the named elements, limitations, steps, integers, and/or features are essential, but other unnamed elements, limitations, steps, integers, and/or features may be added and still form a construct within the scope of the claim. Specific embodiments disclosed herein may be further limited in the claims using the closed-ended transitional phrases“consisting of’ or“consisting essentially of’ (or variations thereof such as “consist of’,“consists of’,“consist essentially of’, and“consists essentially of’) in lieu of or as an amendment for“comprising.” When used in the claims, whether as filed or added per amendment, the closed-ended transitional phrase“consisting of’ excludes any element, limitation, step, integer, or feature not expressly recited in the claims. The closed-ended transitional phrase “consisting essentially of’ limits the scope of a claim to the expressly recited elements, limitations, steps, integers, and/or features and any other elements, limitations, steps, integers, and/or features that do not materially affect the basic and novel characteristic(s) of the claimed subject matter. Thus, the meaning of the open-ended transitional phrase “comprising” is being defined as encompassing all the specifically recited elements, limitations, steps and/or features as well as any optional, additional unspecified ones. The meaning of the closed-ended transitional phrase “consisting of’ is being defined as only including those elements, limitations, steps, integers, and/or features specifically recited in the claim whereas the meaning of the closed-ended transitional phrase“consisting essentially of’ is being defined as only including those elements, limitations, steps, integers, and/or features specifically recited in the claim and those elements, limitations, steps, integers, and/or features that do not materially affect the basic and novel characteristic(s) of the claimed subject matter. Therefore, the open-ended transitional phrase “comprising” (and equivalent open-ended transitional phrases thereof) includes within its meaning, as a limiting case, claimed subject matter specified by the closed-ended transitional phrases“consisting of’ or“consisting essentially of.” As such embodiments described herein or so claimed with the phrase“comprising” are expressly or inherently unambiguously described, enabled and supported herein for the phrases“consisting essentially of’ and“consisting of.”

[086] All patents, patent publications, and other references cited and identified in the present specification are individually and expressly incorporated herein by reference in their entirety for the purpose of describing and disclosing, for example, the compositions and methodologies described in such publications that might be used in connection with the present invention. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard is or should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicant and does not constitute any admission as to the correctness of the dates or contents of these documents.

[087] Lastly, the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Accordingly, the present invention is not limited to that precisely as shown and described.

Claims

2. The lateral flow device of claim 1, wherein the plurality of capture reagent zones further comprise one or more additional capture reagent zones, each of the one or more additional capture zones comprising a competitive ligand, the one or more additional capture zones each serving as an indicator for the presence or absence of the target analyte to the chemosensory protein.

3. The lateral flow device of any one of claim 1 or 2, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain the same amount of the competitive ligand.

4. The lateral flow device of claims 1-3, wherein first capture reagent zone and each of the one or more additional capture reagent zones contain a different amount of the competitive ligand.

5. The lateral flow device of claims 1-4, wherein the different amount of the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones established a concentration gradient.

6. The lateral flow device of any one of claims 1 -5, wherein the matrix further comprises a sample application zone, an absorbent zone, or both a sample application zone and an absorbent zone.

7. The lateral flow device of any one of claims 1-6, wherein the chemosensory protein is an odorant binding protein.

8. The lateral flow device of claim 7, wherein the odorant binding protein is an Anopheles gambiae odorant binding protein 1, an Anopheles gambiae odorant binding protein 32, or an Anopheles gambiae odorant binding protein 33.

9. The lateral flow device of claim 7 or 8, wherein the odorant binding protein is SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4 or an amino acid sequence having at least 85%, at least 90%, at least 95%, at least 97%, at least 98% or at least 99% identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, or SEQ ID NO: 4.

10. The lateral flow device of any one of claim 1-9, wherein nanoparticle is a colloidal gold particle, a colored latex particle, a carbon particle, a selenium particle, a chemiluminescent particle, a bioluminescent particle, a fluorescent particle, a quantum dot particle, an upconverting phosphor particle, a liposome including a dye.

11. The lateral flow device of any one of claims 1-10, wherein nanoparticle is about 15 nmto about 800 nm in size.

12. The lateral flow device of any one of claims 1-11, wherein the competitive ligand present in the first capture reagent zone and each of the one or more additional capture reagent zones is conjugated to a carrier protein.

13. The lateral flow device of claim 12, wherein the carrier protein is a serum albumin or a thyroglobin.

14. The lateral flow device of any one of claims 1-13, wherein the competitive ligand is indole, 3- methyl indole or another derivative of indole.

15. The lateral flow device of any one of claims 1-14, wherein the sample application zone, the competitive reagent zone, the detection zone, and the absorbent zone are each composed of a material that is the same.

16. The lateral flow device of claim 15, wherein the material is nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

17. The lateral flow device of any one of claims 1-14, wherein the sample application zone, the competitive reagent zone, the absorbent zone, or any combination thereof is composed of a material that is different from the material comprising the detection zone.

18. The lateral flow device of claim 17, wherein the sample application zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon.

19. The lateral flow device of claim 17, wherein the competitive reagent zone is composed of cellulose fibers, cross-linked silica, glass microfiber, polyester, or rayon.

20. The lateral flow device of claim 17, wherein the absorbent zone is composed of cotton fibers or cellulose fibers.

21. The lateral flow device of claim 17, wherein the detection zone is composed of nitrocellulose, nylon, polyethersulfone, polyethylenemylar, or plastic-cast membranes having a capillary rise of between 75 sec/4 cm to 240 sec/4 cm.

22. The lateral flow device of any one of claims 1-21, wherein the device further comprises a solid support.

23. The lateral flow device of claim 22, wherein the solid support is a laminate backing material mylar or polyester.

24. A testing device for determining the presence or absence of a target analyte in a liquid sample, the testing device comprising: a) a lateral flow device as defined in claims 1-23; b) a housing that encloses the matrix.

25. The testing device of claim 24, wherein the housing further comprises a sample port and viewing port.

26. A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in claims 1-23 or a testing device of claim 24 or 25; and b) a sample container comprising a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or a derivative thereof.

27. A testing kit for determining the presence or absence of a target analyte in a liquid sample, the testing kit comprising: a) a lateral flow device as defined in claims 1-23 or a testing device of claim 24 or 25; and b) a sample container.

28. The testing kit of claim 26 or 27, wherein the kit further includes instructions on how to perform a method of determining the presence or absence of a target analyte in a liquid sample.

29. The testing kit of any one of claims 26-28, wherein the chemosensory protein conjugate is lyophilized or freeze-dried.

30. The testing kit of any one of claims 26-29, wherein the sample container further comprises a culture media capable of supporting bacterial growth.

31. A testing container for determining the presence or absence of a target analyte in a liquid sample, the testing container comprising: a) a first compartment comprising a lateral flow device as defined in claims 1-23 or a testing device of claim 24 or 25; b) a second compartment comprising a sample region; and c) a partition comprising a seal composed of a breakable material covering a channel, wherein the first compartment and the second compartment are separated by the partition; wherein when the seal covering the channel is intact, the seal prevents fluid communication between the first and second compartments via the channel; and wherein when the seal covering the channel is broken, fluid communication between the first and second compartments via the channel can occur.

32. The testing container of claim 31, wherein the sample region includes a culture media capable of supporting bacterial growth.

33. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) contacting a liquid sample to a lateral flow device as defined in any one of claims 1-23, a testing device according to claim 24 or 25, a testing kit as defined in any one of claims 26- 30, or a testing container as defined in claims 31 or 32; b) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and c) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

34. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) mixing a liquid sample with a chemosensory protein conjugate, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; b) contacting the liquid sample to a lateral flow device as defined in any one of claims 1-23, a testing device according to claim 24 or 25, a testing kit as defined in any one of claims 26-30, or a testing container as defined in claims 31 or 32; c) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and d) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

35. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) contacting the sample-media mixture to a lateral flow device as defined in any one of claims 1-23, a testing device according to claim 24 or 25, a testing kit as defined in any one of claims 26-30, or a testing container as defined in claims 31 or 32; d) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and e) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

36. A method of determining the presence or absence of a target analyte in a liquid sample, the method comprising the steps of: a) adding a liquid sample to a culture media capable of supporting bacterial growth to form a sample-media mixture; b) incubating the sample-media mixture for a period of time; c) adding a chemosensory protein conjugate to the sample-media mixture to form a conjugate mixture, the chemosensory protein conjugate comprising a chemosensory protein covalently linked to a nanoparticle and a target analyte or derivative thereof; d) contacting the conjugate mixture to a lateral flow device as defined in any one of claims 1- 23, a testing device according to claim 24 or 25, a testing kit as defined in any one of claims 26-30, or a testing container as defined in claims 31 or 32; e) permitting the liquid sample to flow through the competitive reagent zone and the detection zone in a manner that allows the chemosensory protein conjugate to interact with the competitive ligand present in the first capture zone and/or the one or more additional capture reagent zones and the antibody having specificity for the chemosensory protein present in the second capture reagent zone; and f) observing the detection zone to determine the presence or absence of analyte in the liquid sample, wherein visualization of only the second capture reagent zone is indicative of the absence of target analyte in the liquid sample; and wherein visualization of at least one of the first capture zone and/or the one or more additional capture reagent zones is indicative of the presence of target analyte in the liquid sample.

37. The method of claim 35 or 36, wherein the period of time in step (b) is at most 1 hour, at most 2 hours, at most 3 hours, at most 4 hours, at most 6 hours., or at most 8 hours.

38. The method of any one of claims 32-36, wherein the method can detect the presence of at least 1 ppm of target analyte, at least 10 ppm of target analyte, or at least 100 ppm of target analyte.

39. The method of any one of claims 32-37, wherein the liquid sample is a water sample.