Method and compositions for isolation, diagnosis and treatment of polyanion-binding microorganisms

• US 6,197,568 B1
• Filed: 07/28/1998
• Issued: 03/06/2001
• Est. Priority Date: 07/29/1997
• Status: Expired due to Fees

• Current Assignee: Health Research Council of New Zealand (Government of New Zealand), Iowa State University Research Foundation Incorporated (Iowa State University), Board of Regents of the University of Michigan (University of Michigan)
• Original Assignee: Board of Regents of the University of Michigan (University of Michigan)
• Inventors: Chen, Yaping, Marks, Rory M., Maguire, Terence, Linhardt, Robert J.
• Primary Examiner(s): Allen, Marianne P.
• Assistant Examiner(s): ZEMAN, MARY K
• Application Number:
  US09/123,770
• Time in Patent Office: 952 Days
• Field of Search: 435/7.1, 435/5, 435/7.8, 435/29, 435/239, 435/803, 435/948, 424/185.1, 424/196.11, 424/197.11, 424/193.1, 424/279.1, 514/23, 514/54-62
• US Class Current: 435/239
• CPC Class Codes: C12N 15/85 for animal cells
• CPC Class Codes: C12N 2770/24151 Methods of production or purification of viral material
• CPC Class Codes: C12N 2810/60 from viruses
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00
• CPC Class Codes: C12Q 1/70 involving virus or bacteriophage immunoassay for viruses G01N33/56983
• CPC Class Codes: G01N 33/56983 Viruses
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
• CPC Class Codes: Y10S 435/803 Physical recovery methods, e.g. chromatography, grinding
• CPC Class Codes: Y10S 435/948 using viruses or cell lines

Abstract

Methods and compositions for the isolation, diagnosis and treatment of microorganisms such as flaviviruses and other hemorrhagic fever viruses are based on the sulfated polyanion-dependent interaction of flaviviruses and hemorrhagic fever viruses, in particular dengue virus, with target cells. The cellular receptors targeted by these viruses have been identified as sulfated polyanionic glycoproteins, that include highly sulfated heparan sulfate glycosaminoglycans for some target cell types, and as a sulfated mucin on vascular endothelium. Compounds such as heparin, highly sulfated heparan sulfate, and synthetic polyanions such as Suramin, inhibit the interaction between the microorganisms and target cells, thereby disrupting the infective process.

Claims

1. A method for detecting a polyanion-binding microorganism in a biological sample, said method comprising:

a) contacting the biological sample with an immobilized polyanion to which the microorganism from said biological sample adheres to form a polyanion-microorganism complex while the remainder of the biological sample is non-adherent to said polyanion;

b) analyzing the complex to determine whether the microorganism is present; and

c) releasing intact microorganism from said complex.

2. The method of claim 1, wherein the microorganism is a virus.

3. The method of claim 2, wherein the virus is a hemorrhagic fever virus.

4. The method of claim 2, wherein the virus is Hepatitis C.

5. The method of claim 2, wherein the virus is a flavivirus.

6. The method of claim 5, wherein the virus is a dengue virus.

7. The method of claim 6, wherein the dengue virus binds the polyanion through a polyanion binding motif having the sequence of SEQ ID NO:

1.

8. The method of claim 6, wherein the dengue virus binds the polyanion through a polyanion binding motif having the sequence of SEQ ID NO:

2.

9. The method of claim 1, wherein the polyanion is selected from the group consisting of heparin, highly sulfated heparan sulfate, a synthetic polyanion, and a dengue virus envelope protein receptor derived from Vero cell glycosaminoglycan (GAG) or endothelial mucin or analogues thereof.

10. The method of claim 1, wherein said immobilized polyanion is a sulfated heparin-derived decasaccharide.