Antibody detection method and device for a saliva sample from a non-human animal

A lateral flow immunoassay (LFIA) apparatus for detection of an analyte in a sample of saliva from a dog, wherein exposure to a virus selected from a group consisting of canine parvovirus (CPV) and canine distemper virus (CDV) is detected, utilizing a test cassette, comprised of:

• a) a test cassette housing;
• b) a sample port formed in said test cassette housing for accessing a sample pad;
• c) said sample pad accessed via said sample port for receiving the saliva sample, wherein said sample pad filters out test-interfering glycoproteins and interfering antibodies and wherein said sample pad contains a polyclonal antibody (Pab) selected from the group consisting of Pab anti-IgG and Pab anti-IgA;
• d) a strip of material that allows lateral flow of liquid and is fitted within said cassette housing, wherein said strip of material is in contact with said sample pad, wherein said strip has an upstream proximal end and a downstream distal end;
• e) a first permeable conjugate pad in contact with said strip, wherein said first permeable conjugate pad contains a species selected from the group consisting of unfixed ligand to which the analyte will bind and antibody proteins that compete with the analyte for a ligand, wherein said first pad species is coupled with biotin to produce a first conjugate;
• f) a second permeable conjugate pad is downstream of said first permeable conjugate pad and in contact with said lateral flow strip, wherein said second permeable conjugate pad contains an unfixed-to-strip ligand that does not bind to the analyte and is avidin bound to a detectable label species thereby forming a second conjugate, wherein said first conjugate encounters and binds to said second conjugate to form a third conjugate;
• g) a first tracer affixed to said strip in an analyte detection zone that is downstream from said second permeable conjugate pad, wherein said tracer forms a detectable complex producing a visible mark upon encountering said third conjugate in said analyte detection zone;
• h) a control zone downstream of said analyte detection zone, wherein said control zone comprises a bound a second tracer directed toward said second permeable conjugate pad unfixed-to-strip ligand and forms a visible complex upon binding;
• and
• i) an absorbent pad that is in contact with the distal end of said strip for absorbing liquid from said strip.

A method utilizing lateral flow immunoassay (LIFA) for detecting an analyte in a body fluid sample from an animal pet, comprising the steps of:

• a) providing a test cassette, wherein said test cassette comprises:
• i) a test cassette housing;
• ii) a sample port formed in said test housing that accesses a sample pad for receiving the body fluid sample;
• iii) a strip of material that allows lateral flow of liquid and is fitted within said cassette housing, wherein said strip of material is in contact with said sample pad;
• iv) a first permeable conjugate pad in contact with said strip, wherein said first permeable conjugate pad contains a species selected from a group consisting of unfixed ligand to which the analyte will bind and antibody proteins that compete with the analyte for a ligand, wherein said first pad species is coupled with biotin to produce a first conjugate;
• v) a second permeable conjugate pad is downstream of said first permeable conjugate pad and in contact with said lateral flow strip, wherein said second permeable conjugate pad contains an unfixed ligand that does not bind to the analyte and is unbound avidin bound to a detectable label species thereby forming a second conjugate, wherein said first conjugate encounters and binds to said second conjugate to form a third conjugate;
• vi) a first tracer affixed to said strip in an analyte detection zone that is downstream from said second permeable conjugate pad, wherein said tracer forms a detectable complex producing a visible mark upon encountering said third conjugate in said analyte detection zone;
• vii) a control zone downstream of said analyte detection zone, wherein said control zone comprises a second bound tracer directed toward said second permeable conjugate pad unfixed ligand and forms a visible complex upon binding;
• and
• viii) an absorbent pad that is in contact with the distal end of said strip for absorbing liquid from said strip;
• b) applying the sample via said sample port to said test cassette;
• and
• c) detecting a visual indicator to establish a presence of the analyte.

A method utilizing lateral flow immunoassay (LIFA) for detecting of an analyte in a sample of saliva from a dog, wherein exposure to a virus selected from a group consisting of canine parvovirus (CPV) and canine distemper virus (CDV) is detected, comprising the steps of:

• a) providing a test cassette, wherein said test cassette comprises:
• i) a test cassette housing;
• ii) a sample port formed in said test housing that accesses a sample pad for receiving the body fluid sample;
• iii) a strip of material that allows lateral flow of liquid and is fitted within said cassette housing, wherein said strip of material is in contact with said sample pad;
• iv) a first permeable conjugate pad in contact with said strip, wherein said first permeable conjugate pad contains a species selected from a group consisting of unfixed ligand to which the analyte will bind and antibody proteins that compete with the analyte for a ligand, wherein said first pad species is coupled with biotin to produce a first conjugate;
• v) a second permeable conjugate pad is downstream of said first permeable conjugate pad and in contact with said lateral flow strip, wherein said second permeable conjugate pad contains an unfixed ligand that does not bind to the analyte and is unbound avidin bound to a detectable label species thereby forming a second conjugate, wherein said first conjugate encounters and binds to said second conjugate to form a third conjugate;
• vi) a first tracer affixed to said strip in an analyte detection zone that is downstream from said second permeable conjugate pad, wherein said tracer forms a detectable complex producing a visible mark upon encountering said third conjugate in said analyte detection zone;
• vii) a control zone downstream of said analyte detection zone, wherein said control zone comprises bound a second tracer directed toward said second permeable conjugate pad unfixed ligand and forms a visible complex upon binding;
• and
• viii) an absorbent pad that is in contact with the distal end of said strip for absorbing liquid from said strip;
• b) applying the sample via said sample port to said test cassette;
• and
• c) detecting a visual indicator to establish a presence of the analyte.