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Diagnosis and management of infection caused by Chlamydia

a technology of chlamydia and chlamydia spores, applied in the direction of biocide, heterocyclic compound active ingredients, drug compositions, etc., can solve the problems of c. pneumoniae /i>infections may relapse, chlamydia is difficult to eradicate, etc., to prevent chlamydia from reactivation and the effect of preventing the reactivation of the pathogen

Inactive Publication Date: 2005-06-30
VANDERBILT UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention provides a unique approach for the diagnosis and management of infection by Chlamydia species, particularly C. pneumoniae. The invention involves the use of a combination of antichlamydial agents, each targeted against a different phase of the chlamydial life cycle. The agents can be administered simultaneously or sequentially, and can include anti-inflammatory agents, immunosuppressive agents, and anti-porphyrial agents. The invention also provides a method for evaluating the infection status of an individual and monitoring the progress of therapy for treating Chlamydia infection. The invention can help to significantly reduce or eliminate chlamydial infection, and can be used in the treatment of diseases associated with Chlamydia infection. The invention also provides a susceptibility test for identifying agents capable of significantly reducing or eliminating chlamydial infection.

Problems solved by technology

Once fully established, the Chlamydia are difficult to eradicate, with frequent relapse following antibiotic therapy.
Despite this demonstration of in vitro susceptibility, C. pneumoniae infections may relapse following antibiotic therapy with these agents.

Method used

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  • Diagnosis and management of infection caused by Chlamydia
  • Diagnosis and management of infection caused by Chlamydia
  • Diagnosis and management of infection caused by Chlamydia

Examples

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example 1

POLYMERASE CHAIN REACTION (PCR) FOR THE FULL LENGTH MOMP GENE OF C. PNEUMONIAE AND OTHER SPECIES OF CHLAMYDIA (DIAGNOSTIC)

[0200] a. Solution PCR

[0201] Serum, blood or tissue samples were pre-incubated in the presence of 10 μM dithiothreitol at room temperature for 2 hours to reduce the disulfide bonds and facilitate release of the outer shell of the elementary bodies. CSF and other body fluids are also suitable for use as described. Other suitable reducing agents for use in this step include, but are not limited to, succimer and glutathione (e.g., including, but not limited to, glutathione esters, other analogs and deriviatives). The failure to include a reducing agent initially may result in a negative PCR signal following the protease digestion step. Appropriate concentrations of these reducing agents can be readily determined by the skilled artisan without undue experimentation using the 10 μM concentration of dithiothreitol as a guideline. Alternatively, guanidine isothiocyana...

example 2

ENZYME LINKED IMMUNO SORBENT ASSAY (ELISA; DIAGNOSTIC)

[0206] a. Recombinant MOMP-Based ELISA

[0207] The full length MOMP gene of C. pneumoniae was directionally cloned into the pET expression plasmid at the NCOI and NOTI restriction sites using primers to introduce these unique restriction sites into the MOMP ends. Primer sequences are as follows:

CPOMPDNCO (Coding strand):(SEQ ID NO. 43)5′-AGCTTACCAT GGCTAAAAAA CTCTTAAAGT CGGCGTTATTATCCG-3′CPOMP_CNOT (complimentary strand):(SEQ ID NO. 44)5′-ATATGCGGCC GCTCATAGAA TCTGAACTGA CCAGATACG-3′

[0208] The construction of the MOMP insert into the pET expression vector (Novagen, Inc.) yields, on transformation of permissive E. coli, an amino terminal thioredoxin fusion domain, a polyhistidine for Ni+-affinity chromatography, a solubility sequence of approximately 5 kD, and an endopeptidase cleavage site which yields a full length MOMP with a modified amino terminal (as illustrated in FIG. 2) containing an alanine insert between the amino ter...

example 3

DETECTION ASSAY METHODS (DIAGNOSTIC)

[0212] a. Immunoglobulin (Ig) assay

[0213]C. pneumoniae EBs were grown in primary human umbilical vein endothelial cells (HuEVEC; early passage), HeLa 199, or a suitable alternative in the presence of 1 μg / ml cycloheximide at 35° C. under 5% CO2. Permissive cells were lysed by sonication at 3 days, thereby liberating EBs. The latter were harvested from infection flasks, sonicated, and cellular debris were removed after sonication by a low speed centrifugation (˜600×g) for 5 minutes. EBs were pelleted by high speed centrifugation (30,000×g) for 30 minutes at 4° C. The EB pellet was washed with PBS ×1 and was reconstituted in 2 ml PBS per four 25-cm2 culture flask and sonicated at maximum power for 20 seconds and a 0.5 cycle time using a Braun-Sonic U sonicator. EB protein concentration was determined by the Bradford method and the sonicated infectious EB suspension was rendered non-infectious by the addition of 37% formaldehyde to a final 10% form...

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PUM

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Abstract

The present invention provides a method of treating coronary artery disease in a patient in need thereof by administering to the patient an antichlamydial amount of a rifamycin for a duration to treat said coronary artery disease.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application is a continuation application of Ser. No. 10 / 100,759, filed Mar. 19, 2002, which is a continuation application of and claims priority to U.S. Ser. No. 09 / 073,661, filed May 6, 1998, which is a continuation-in-part of U.S. Ser. No. 09 / 025,521, filed Feb. 18, 1998, which is a continuation-in-part of U.S. Ser. No. 08 / 911,593, filed Aug. 14, 1997. U.S. Ser. No. 09 / 073,661 is also a continuation-in-part of U.S. Ser. No. 09 / 025,176 and U.S. Ser. No. 09 / 025,174, each filed Feb. 18, 1998, and claims benefit of U.S. Provisional Application Nos. 60 / 045,739, 60 / 045,779, 60 / 045,780, 60 / 045,784, 60 / 045,787, and 60 / 045,689, each filed May 6, 1997. Each of the foregoing applications is incorporated herein by reference.BACKGROUND OF THE INVENTION [0002]Chlamydiae are obligate intracellular microorganisms which parasitize eukaryotic cells and are ubiquitous throughout the animal kingdom. Members of the chlamydial genus are considered ba...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K31/455A61K45/06
CPCA61K31/192A61K31/455A61K31/496A61K31/573Y10S514/824A61K45/06A61K31/7052A61K2300/00
Inventor MITCHELL, WILLIAMSTRATTON, CHARLES
Owner VANDERBILT UNIV
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