Solid phase test device for sialidase assay

• US 7,591,978 B2
• Filed: 08/10/2006
• Issued: 09/22/2009
• Est. Priority Date: 08/10/2006
• Status: Expired due to Fees

• Current Assignee: Alere Switzerland GmbH (Abbott Laboratories Incorporated)
• Original Assignee: Inverness Medical Switzerland GmbH (Abbott Laboratories Incorporated)
• Inventors: Fish, Falk, Keren, Tomer, Dwir, Oren Shraga
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US11/502,192
• Publication Number:
  US 20080038766A1
• Time in Patent Office: 1,139 Days
• Field of Search: 435/25, 435/18, 422/56
• US Class Current: 422/412
• CPC Class Codes: C12Q 1/34 involving hydrolase
• CPC Class Codes: C12Q 2334/50 Indoles
• CPC Class Codes: C12Q 2334/52 5-Bromo-4-chloro-3-indolyl, i.e. BCI
• CPC Class Codes: G01N 2333/195 from bacteria
• CPC Class Codes: G01N 2333/30 from Mycoplasmatales, e.g. Pleuropneumonia-like organisms [PPLO]
• CPC Class Codes: G01N 2333/36 from Actinomyces; from Streptomyces (G)
• CPC Class Codes: G01N 2800/36 Gynecology or obstetrics

Abstract

A solid-phase test device and a method for the detection of sialidase activity in a fluid sample. The test device includes a solid support provided with a pre-deposited dry form of a sialidase detecting composition. The sialidase detecting composition includes a sialidase substrate and a color-developing reagent. When the sialidase substrate is exposed to sialidase it yields an intermediate compound which reacts with the color-developing reagent to form a detectable color change.

Claims

1. A solid-phase test device for the detection of sialidase activity in a fluid sample, configured as a strip for a lateral flow assay, the device comprising a first flow matrix, a second flow matrix and an absorbent pad sequentially arranged on a non-absorbing solid support in fluid communication with each other to allow flow of fluid from said first flow matrix through said second flow matrix into said absorbent pad, wherein a sialidase detecting composition comprising a sialidase substrate and a color-developing reagent is movably bound in a re-suspendable form on said first flow matrix, and wherein said sialidase substrate when exposed to sialidase yields an intermediate compound which reacts with said color-developing reagent to form a detectable color change.

2. The test device of claim 1 wherein said first flow matrix and second flow matrix are of different permeation rates, the permeation rate through the first flow matrix being higher than the permeation rate through the second flow matrix.

3. The test device of claim 1 wherein said first flow matrix is a glass fiber matrix or a filter paper.

4. The test device of claim 1 wherein said second flow matrix is a nitrocellulose membrane or a nylon membrane.

5. The test device of claim 1 wherein said first flow matrix and second flow matrix are arranged so as to form an overlapping interface zone therebetween.

6. The test device of claim 5 wherein said overlapping interface zone is of about 1 to 5 mm length in the direction of liquid transport.

7. The test device of claim 1 wherein the first flow matrix is a glass fiber or a polymeric membrane and said second flow matrix is a nitrocellulose or nylon membrane.

8. The test device of claim 1 wherein at least one product of the reaction between said intermediate compound and said color-developing reagent is an insoluble color precipitate.

9. The test device according to claim 8 wherein said precipitate is accumulated at an interface between the first and second flow matrices.

10. The test device of claim 1 wherein the first flow matrix comprises a first zone impregnated with the sialidase substrate, a second zone impregnated with the color-developing reagent and a third zone for receiving a sample.

11. The test device of claim 10 wherein said first, second and third zones are overlapping.

12. The test device of claim 10 wherein said third zone is located upstream of said first and second zones.

13. The test device of claim 10 wherein said first zone is located upstream of said second zone.

14. The test device of claim 1 wherein said fluid sample is a vaginal discharge sample.

15. The test device of claim 1 wherein the detectable color change forms a signal line at an interface between the first and the second flow matrices.

16. The test device of claim 1 wherein the reaction between said intermediate compound and said color-developing regent is an oxidation-reduction reaction.

17. The test device of claim 1 wherein said sialidase substrate comprises a sialic moiety linked to a non-sialic moiety by a ketoside bond and wherein said non-sialic moiety is an indoxyl or a derivative thereof.

18. The test device of claim 17 wherein said sialic moiety is N-acetylneuraminic acid or a derivative thereof.

19. The test device of claim 1 wherein said sialidase substrate is 5-bromo-4-chloro-3-indolyl-α

-D-N-acetylneuraminic acid or a salt thereof.

20. The test device of claim 1 wherein said composition further comprises a surfactant.