TH18160Y - A colorimetric test kit for detecting type B toxin that causes early mortality syndrome (EMS) in shrimp. - Google Patents

Abstract

------10/09/2563------(OCR) This invention describes a ready-to-use test kit for detecting type B toxin causing early mortality syndrome (EMS) in shrimp. It consists of a rectangular cassette containing a nitrocellulose membrane with a test line containing MAB. The kit includes an EMSB-39B (MAb EMSB-39B) line and a control line containing a goat anti-mouse IgG antibody (GAM). A glass fiber pad containing colloidal gold (dark purple) is connected to an EMSB-5B specific toxin B that causes early mortality syndrome (EMS). A sample pad and an absorbing pad are also included. This colorimetric EMS test kit diagnoses toxin B produced by EMS bacteria. When a sample is placed in the kit, the toxin B is absorbed by the gold-connected EMSB-5B, flows through the nitrocellulose pad, and is absorbed again by the EMSB. EMSB-39B (MAb EMSB-39B) is drawn on the nitrocellulose plate at the T region in a sandwich-like manner, forming a colored band where the MAb is drawn at the T region. Conversely, MAb EMSB-5B, connected to a gold colloid that either binds excess or non-binding toxin B, flows through the T region and is captured by goat anti-mouse IgG antibody (GAM), forming a colored band at the C region. For minced shrimp samples without toxin or with toxin levels below the kit's sensitivity, only a single colored band appears at the C region. This kit does not cross-react with other shrimp pathogens and is a unique invention unlike any other. It is easy to use, can be administered by the general public without requiring specialized knowledge, additional equipment, or laboratory facilities, and is suitable for field use. Provides accurate, precise, and rapid results within 15 minutes. ------------ Page 1 of 1. Summary of the Invention: The colorimetric strip test kit for detecting early mortality syndrome (EMS) in shrimp consists of two types of magnetic reactive organisms (MAbs) that specifically bind to toxin 5B produced by pathogenic bacteria. One MAb, EMSB-5B, is connected to a dark purple gold colloid, and the other MAb, EMSB-39B, is marked on a porous nitrocellulose plate at the T region. The C region is marked with GAM. When a sample is placed in the test kit, the toxin B is bound by the first MAb connected to the gold colloid, flows through the nitrocellulose plate, and is bound again by the MAB marked on the nitrocellulose plate (T region) in a sandwich-like manner. A colored band appears where the MAB is marked (region T). The EMSB-5B MAB, which contains gold colloid but is not reacting with the toxin, flows through the T band and is reacted with by GAM, forming a colored band at region C. For minced shrimp samples without toxin or with toxin levels lower than the kit's sensitivity, only a single colored band will appear at region C. This is a ready-to-use kit that is easy to use and can be administered by the general public without the need for additional equipment or laboratory facilities.

Claims (2)

------10/09/2563------(OCR) 1. A colorimetric test kit for early mortality syndrome (EMS) in shrimp, which consists of a backing plate (1) which is a plastic sheet for attaching other components of the colorimetric test kit for early mortality syndrome (EMS) in shrimp, and a nitrocellulose membrane (2) which consists of two lines: one test line (3) and one control line (4), where the test line (3) is the line containing the MAB. EMSB-39B (MAb EMSB-39B) specific toxin type B causing early death is drawn on the nitrocellulose pad at T. The control line (4) is a line with goat anti-mouse IgG antibody (GAM) specific toxin drawn on the nitrocellulose pad at C. The glass fiber pad (5) is a pad with dark purple colloidal gold connected to the MAb. The EMSB-5B (MAb EMSB-5B) test strip, which is specific to the early-onset b-toxin, is placed on top of a glass fiber filter (5) on a nitrocellulose sheet (2). The sample pad (6) is the part that absorbs the sample to diffuse and pass to the glass fiber filter (5), which is placed under the sample pad (6). The absorbing pad (7) is the part that absorbs the remaining sample and is placed on the nitrocellulose sheet (2) opposite the sample pad (6). The test cassette is a rectangular cassette with two flaps that fit together. The outer top flap has an oval sample receiving opening (8) and is printed with the letters T (3) and C (4) on the outside. The other side is printed with the letters EMSB (9). The inner top flap has a ridge (10) that holds the EMSB-5B test strip in place. On the inside of the lower lid, there is a ridge (11) which is a base for placing the Early Mortality Syndrome (EMS) test strip, which is attached to the lower lid. A sealed, moisture-free aluminum foil pouch is used to package the Early Mortality Syndrome (EMS) test strip kit. 2. Early Mortality Syndrome (EMS) test strip kit according to claim 1, where the pad (1) is a non-absorbent plastic sheet and does not react with the sample and antigens or antibodies contained in the Early Mortality Syndrome (EMS) test strip. 3. Early Mortality Syndrome (EMS) test strip kit according to claim 1 or 2, where the pad (1) has adhesive strips that divide it into sections for attaching other components, namely a nitrocellulose sheet. (nitrocellulose membrane) (2) glass fiber pad (5) sample pad (6) and absorbing pad (7) 4. A color strip test kit for early mortality syndrome (EMS) in shrimp according to claim 1, in which the test line (3) is a line containing MAb EMSB-39B which is specific to early mortality syndrome (EMS) toxin at a concentration of 1-2 mg. 5. A color strip test kit for early mortality syndrome (EMS) in shrimp according to claim 1 or 4, in which the test line (3) is a line containing MAb EMSB-39B which is specific to early mortality syndrome (EMS) toxin at a concentration of 1-2 mg. 6. A color strip test kit for early mortality syndrome (EMS) in shrimp, according to claim 1, where the control line (4) is a line containing goat anti-mouse IgG antibody or GAM with a concentration of not less than 1 mg. 7. A color strip test kit for early mortality syndrome (EMS) in shrimp, according to claim 1 or 6, where the control line (4) is a line containing goat anti-mouse IgG antibody or GAM with an optimal concentration of 0.8 mg. 8. A color strip test kit for early mortality syndrome (EMS) in shrimp, according to claim 1, where the fiberglass plate (5) is a plate containing colloidal gold. 9. Early mortality syndrome (EMS) color strip test kit according to claim 1 or 8, in which the glass fiber plate (5) is a plate with colloidal gold, dark purple in color, attached to MAb EMSB-5B, which is specific to early mortality syndrome (EMS) toxin, at a concentration of 1-2 mg/ml. The optimal concentration is 1 milligram per milliliter (mg/ml). 10. A colorimetric test kit for early mortality syndrome (EMS) in shrimp, as per claim 1, in which a rectangular cassette with two tightly fitting lids holds the assembled colorimetric test kit for early mortality syndrome (EMS). ------------ Page 1 of 1 page Claims 1. The color strip test kit for early mortality syndrome (EMS) in shrimp consists of a rectangular cassette (1) with two tightly fitting lids. The upper lid (A) has an oval sample receiving slot and the letters T and C at the position of the MAb (EMSB-39B) and GAM (GAM) marks, respectively. The letters EMSB are printed on the sample receiving plate on the lower lid (B). The lower lid has a ridge in the middle for inserting the test kit, which is attached to the lower lid. The other MAb, EMSB-5B, is connected to a dark purple colloidal gold placed on a fiberglass plate in front of the letter T. A special feature is that both MABs (EMSB-5B and EMSB-39B) are strong and specific receptors for type B toxin. 2. Pursuant to claim 1, one type of MAB (EMSB-5B) is bonded to a dark purple gold colloid and another type of MAB (EMSB-39B) is drawn onto a nitrocellulose porous material at region T, and a GAM is drawn at region C.