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Method for detecting syphilis using synthetic antigens

An antigen and composition technology, applied in the fields of microbiology and immunology, can solve the problems of time-consuming, expensive and other problems that cannot be used as a test to detect the success of antibiotic treatment

Inactive Publication Date: 2002-06-12
UNITED STATES OF AMERICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although treponemal-based assays can be used to confirm positive assay results, these assays are often expensive, complex, and time-consuming, and may require the use of highly sophisticated scientific equipment and trained technicians
Furthermore, the treponemal assay cannot be used as a test to detect the success of antibiotic therapy because anti-Treponema pallidum antibodies persist after cure, and in approximately 85% of successfully treated individuals, the test results remain even after eradication of the infection positive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Embodiment 1 prepares synthetic cardiolipin and lecithin composition

[0059] Tetramyristoyl cardiolipin purified by silica gel chromatography to approximately 99% purity was obtained from Avanti Polar Lipids (Alabaster, AL) in powder form. Final concentrations of sodium salt solutions were tested for purity by TLC and HPLC. Samples were stored at -20°C. The tetramyristoyl cardiolipin was originally synthesized from semi-synthetic lipid precursors of plant origin.

[0060] Lecithin (1-palmitoyl-2-oleoyl-sn-glycerol-phosphocholine) was also obtained from Avanti Polar Lipids, purified by silica gel chromatography to about 99% purity. The lecithin was originally isolated from soybean.

[0061] A 1.2% solution of cholesterol (Avanti Polar Lipids) in absolute ethanol was prepared and filtered through alcohol rinsed filter paper #560. The cholesterol was originally obtained from lanolin and purified by recrystallization, and the crystals were stored at -20°C.

[0062] Sy...

Embodiment 2

[0063] The comparative analysis of the synthetic VDRL slide measurement and conventional VDRL slide measurement of embodiment 2

[0064] The sensitivity of the VDRL slide assay using the synthetic cardiolipin and lecithin compositions described in Example 1 was compared with A Manual of Tests for Syphilis (9th Edition, 159-77, Larsen, S.A., Pope, V., Johnson, R.E. and Kennedy, E.J.Jr. (eds.), American Public Health Association, Washington, DC) compared the sensitivity of conventional VDRL slide assays. Briefly, 0.4 ml of VDRL buffered saline (formaldehyde, Na 2 HPO 4 、KH 2 PO 4 , NaCl and distilled water) were added to the bottom of a 30 ml round bottom flask with a glass stopper or a 25 ml Erlenmeyer flask with a flat inner bottom surface. Then, 0.5 ml of the antigen composition suspension was directly added to the saline solution at a speed of 6 seconds per 0.5 ml of the antigen suspension while continuously rotating the flask. Rotation was continued for 10 seconds unti...

Embodiment 6

[0085] Comparative Analysis of Synthetic and Natural VDRL Antigens in Biological False Positive Samples (Qualitative Test)

[0086] Samples were obtained from 50 individuals originally classified as Biological False Positives (BFPs). These individuals were reactive in the non-treponemal test and non-reactive in the treponemal test. These samples were tested using the procedure of Example 3. The results of this test are listed in Table 7. Four serum samples initially misclassified as BFP were found to be reactive when assayed using CDC synthetic VDRL antigen, TP-PA and FTA-ABS. Three of these four samples were also reactive with native VDRL antigens.

[0087] Sample reactivity

[0088] R = Reactive; N = Non-reactive

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Abstract

An antigen composition and method for the detection of antibodies to Treponema pallidum and the diagnosis of syphilis are described. The antigen composition contains synthetic cardiolipin and synthetic lecithin. The antigen composition may additionally contain cholesterol and an alcohol. The antigen composition is useful as an immunoreagent in immunoassays for the detection of antibodies associated with T. pallidum infection. The methods are sensitive and specific for T. pallidum infection.

Description

[0001] This invention was made by a United States government agency, the Centers for Disease Control and Prevention. field of invention [0002] The present invention relates to the fields of microbiology and immunology, and more particularly, to compositions and methods for detecting, diagnosing and monitoring the treatment of syphilis. In particular, the present invention relates to synthetic cardiolipin and lecithin antigen compositions and their use in immunoassays. Background of the invention [0003] Syphilis is a sexually transmitted disease (STD) caused by the bacterium Treponema pallidum. Worldwide, more than 100,000 cases of syphilis in adults are reported annually. The disease can also be transmitted congenitally, infecting 3000 or more infants each year. When antibiotic treatment is not given in the early stages of a blight, the disease can spread throughout the body, often leading to irreversible damage to organs, mental illness, blindness or death. The world...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N33/571G01N33/92
CPCG01N33/92G01N2405/04G01N33/571G01N2333/20
Inventor 维多利亚·波普阿诺德·R·卡斯特罗威廉·E·莫里尔
Owner UNITED STATES OF AMERICA
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