Methods and kits for the detection of an infection in subjects with low specific antibody levels

A specific, subject-based technique, applied to antibodies against and/or breakthrough infection, detection against latent infection, chronic infection, re-infection, capable of addressing below or just at detection levels, hindering detection specificity, etc. question

Inactive Publication Date: 2013-07-24
智能生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Improving the analytical sensitivity of the assay under development will not solve the diagnostic problem as this would also increase the noise level which would hinder the specificity of the assay
Therefore, there is an unmet need for the development of antibody-based assays to be able to detect XMRV and other retroviral and pathogenic infections where infection has a latent period where antibody levels are below or just at detection levels

Method used

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  • Methods and kits for the detection of an infection in subjects with low specific antibody levels
  • Methods and kits for the detection of an infection in subjects with low specific antibody levels
  • Methods and kits for the detection of an infection in subjects with low specific antibody levels

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0169] Example 1: Activators increase seroresponse to xenotropic murine leukemia virus (XMRV) infection Specific antibodies in subjects who should be positive

[0170] Blood from subjects known to be infected with XMRV was collected in anticoagulants, brought to the laboratory, and within 24 hours at room temperature, 1 ml of blood was transferred to tubes containing mitogen. Test the rest of the plasma for XMRV antibodies ( figure 1 , solid line) and remain for any additional future comparison tests. Incubate blood for 3-5 days in tubes containing mitogens (5% CO 2 , 37°C), the treated plasma was then collected and tested for XMRV antibodies using the same kit / assay used to test untreated plasma.

[0171] XMRV antibody concentrations were higher in mitogen-treated plasma than in untreated plasma (Table 1). Therefore, XMRV infection of processed plasma can be diagnosed with higher sensitivity.

[0172] Table 1. In untreated and mitogen-treated blood from XMRV-infecte...

Embodiment 2

[0179] Example 2: Anti-IGD further increases xenotropic murine leukemia virus (XMRV) infection Raised specific antibodies to the activator in seropositive subjects

[0180] Blood samples collected from individuals infected with XMRV whose plasma antibody levels have fallen to at or below assay detection levels (eg, OD readings near the cutoff value). Aliquots of 1 ml of fresh, non-clotted whole blood (or PBMC) were incubated in anti-IgD, anti-IgD and mitogen, or mitogen alone.

[0181] Table 3: After three days of incubation with anti-IgD, mitogen, or anti-IgD and mitogen After that, the OD reading of the anti-XMRV antibody:

[0182] sample#

plasma (unprocessed)

anti-IgD

Anti-IgD and mitogens

mitogen

1

0.110

0.120

0.400

0.300

2

0.200

0.350

0.600

0.550

3

0.101

0.150

0.380

0.200

4

0.090

0.110

0.180

0.140

5

0.087

0.115

0.240

0.170

...

Embodiment 3

[0185] Example 3: Activators Improve Seropositive Hepatitis E Virus (HEV) Infection Specific antibodies in the subject

[0186] Blood collected in anticoagulant from the local population in Beijing was brought to the laboratory (within 24 h at RT) and 1 ml of blood was transferred to a tube containing activator (SMARTube TM , which can be obtained from http: / / www.smartube-bio.com / ?CategoryID=190 , accessed September 6, 2011) in the container. The remainder of the plasma (untreated) was tested for HEV antibodies and kept for any additional future comparative testing. Incubate the blood in the container for 3-5 days (5% CO 2 , 37°C), plasma was then collected and tested for HEV antibodies using the same kit / assay used to test unprocessed blood samples.

[0187] Table 4: HEV antibody levels after stimulation of whole blood with activators (ELISA)

[0188]

[0189] Table 4 shows the results for 7 / 15 seropositive samples in which antibody levels increased after stimul...

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Abstract

This invention relates to methods that enable the detection of antibodies against a latent infection, a chronic infection, a re-infection, and / or a breakthrough infection; enable the diagnosis of a latent infection, a chronic infection, a re-infection, and / or a breakthrough infection; and increase low anti-viral antibody levels, and a kit for the detection of virus-specific antibodies expressed at low levels.

Description

technical field [0001] The present invention relates to the ability to detect antibodies against latent infection, chronic infection, reinfection, and / or breakthrough infection; to enable diagnosis of latent infection, chronic infection, reinfection, and / or breakthrough infection; and to improve low antiviral antibodies level methods, and relates to kits for the detection of virus-specific antibodies expressed at low levels. Background technique [0002] Following chronic infection with certain pathogens, including viral or retroviral pathogens, the levels of antibodies against the pathogen are too low to be conclusively detected or may not be detected at all. This phenomenon has been observed in some lung infections (e.g., pneumonia and tuberculosis), where the pathogen localizes in "inclusion bodies" that hide it from the immune system, making antigenic stimulation too low to remain measurable over the long term or detectable antibody production. In the case of retrovira...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/567C12N5/071
CPCG01N2469/20G01N33/576G01N33/56983G01N2333/15Y02A50/30
Inventor 塔玛·杰胡达-柯亨
Owner 智能生物科技有限公司
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