Continuous stable cell line for identification of infectious African swine fever virus in clinical samples

Patent metadata

Field	Value
Publication number	US-11796548-B2
Application number	US-202117236284-A
Country	US
Kind code	B2
Filing date	Apr 21, 2021
Priority date	Apr 22, 2020
Publication date	Oct 24, 2023
Grant date	Oct 24, 2023

Abstract

Methods are provided herein utilizing a stable cell line capable of efficient infection by African swine fever virus (ASFV) and also provides for the detection of the presence of virus in samples applied to the cells. Detection of the virus by means such as red blood cell rosetting is a surprising result given that the cell line is derived from African green monkeys. This cell line provides a marked improvement over the currently available testing strategies.

First claim

What is claimed is:

1. A method of detecting African Swine Fever Virus (ASFV) in a sample, comprising the steps of:
a. contacting the sample with a culture of Ma-104 cells under conditions that allow for infection of the cells by ASFV;
b. incubating the Ma-104 cell and sample mixture under conditions that allow for ASFV detection; and
c. analyzing the mixture from step b for the presence or absence of replicating ASFV.

2. The method of claim 1 , wherein the ASFV comprises a strain of ASFV that is not attenuated for growth in Ma-104 cells.

3. The method of claim 1 , wherein the ASFV comprises a strain of ASFV that has not been passaged in cell culture prior to contacting with Ma-104 cells.

4. The method of claim 1 , wherein the ASFV comprises the Georgia/2007 strain.

5. The method of claim 1 , wherein the sample comprises a sample from a swine.

6. The method of claim 1 , wherein the analyzing step comprises adding red blood cells to the mixture from step b, and examining the Ma-104 cells for the presence or absence of hemadsorbed red blood cells on the Ma-104 cells.

7. The method of claim 6 , wherein the presence or absence of hemadsorbed red blood cells is determined within 24 hours after the red blood cells are added to the Ma-104 cell and sample mixture.

8. A composition comprising Ma-104 cells, African Swine Fever Virus (ASFV), and red blood cells.

9. The composition of claim 8 , wherein the ASFV comprises the Georgia/2007 strain.

10. The composition of claim 1 , wherein the Ma-104 cells are genetically unmodified.

11. The composition of claim 8 , wherein the Ma-104 cells are genetically unmodified.

Assignees

• Us Agriculture

Inventors

• Gladue Douglas P
• Rai Ayushi
• Borca Manuel V

Classifications

• G01N33/80 Primary

  involving blood groups or blood types {or red blood cells (white blood cells G01N33/56972)} · CPC title

• C12N7/00

  Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00) · CPC title

• C12N2710/12031

  Uses of virus other than therapeutic or vaccine, e.g. disinfectant · CPC title

• G01N2333/01

  DNA viruses · CPC title

• G01N2800/26

  Infectious diseases, e.g. generalised sepsis · CPC title