 Protease-activated receptor PAR4 (ZCHEMR2) |
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| Xylofuranosly-containing nucleoside phosphoramidites and polynucleotides |
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Compositions and methods for sensitive detection of HIV infection and monitoring of drug resistance
| Details |
Inventors: Dong, Jian-yung;
Assignee: MUSC Foundation for Research Development (Charleston, SC)
Primary Examiner: Stucker; Jeffrey
Assistant Examiner: Winkler; Ulrike
Attorney, Agent or Firm: Weitz; David J., Chen; Shirley Wilson Sonsini Goodrich & Rosati
A method is provided for detecting a presence of HIV virus in a sample comprising: taking a culture of recombinant cells which (a) are capable of cell division, (b) express CD4 receptor and one or more additional cell surface receptors necessary to allow the HIV virus to infect, (c) enable the HIV virus to replicate and infect the noninfected cells in the cell culture, and (d) comprise a reporter sequence introduced into the recombinant cells comprising a reporter gene whose expression is regulated by a protein specific to HIV viruses which is expressed from a genome of an HIV virus upon infection of the recombinant cell by the HIV virus; contacting the cell culture with a sample to be analyzed for the presence of HIV virus in the sample; and detecting a change in a level of expression of the reporter gene in cells in the recombinant cell culture. The method can be used to detect the presence of HIV virus in a sample, detect the presence of different strains of HIV virus in a sample, detect HIV drug resistance in a sample, determine what combination of one or more anti-HIV agents would be effective in treating a patient, and screen compositions for anti-HIV activity. |
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DETAILED DESCRIPTION OF THE INVENTION The present invention relates to new and useful methods including methods for detecting HIV, methods for detecting HIV drug resistance, methods for designing patient customized anti-HIV drug cocktail treatments, and methods for screening compositions for anti-HIV activity.
Also provided are novel cell lines which may be used with the methods of the present invention.
The methods of the present invention use cells which (a) are capable of cell division; (b) are permissive to HIV virus; (c) express a reporter gene whose expression is selectively regulated by infection with HIV; and (d) allow viral replication of HIV in infected cells which enables cells within the same cell culture which are initially uninfected to become infected.
One of the advantages provided by the present invention is that the recombinant cells used are capable of cell division.
As a result, it is easy to produce and maintain these cells for performing the various methods of the present invention.
A further advantage provided by the present invention is that the recombinant cells can be infected by multiple different strains of HIV, including wild-type and mutant HIV strains from clinical isolates or laboratory-adapted strains.
As a result, the methods of the present invention have broad applicability to all strains of HIV.
Yet a further advantage provided by the present invention is that infection of the recombinant cells by an HIV virus can be easily monitored and measured.
By using a reporter gene whose expression is regulated by infection with HIV, it is possible to detect HIV infection by simple detection methods, such as colorimetric methods.
By expression of the reporter gene being selectively regulated by infection with HIV, false positive signals, for example due to infection by non-HIV viruses, are reduced.
A further advantage of the present invention is that the recombinant cells not only allow entry and infection of the HIV virus, but also facilitate efficient replication within the recombinant cell and transmission of the mature HIV virion to infect other cells in the culture
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Gene Therapy 
