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Novel diagnostic marker, a diagnostic kit and a method for diagnosis of rheumatoid arthritis

a diagnostic kit and rheumatoid arthritis technology, applied in the field of new diagnostic markers, can solve the problems of apf not being widely used, immunofluorescence not becoming popular in clinical practice, and hampered diagnosis of rheumatoid arthritis

Inactive Publication Date: 2006-06-29
COUNCIL OF SCI & IND RES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The present invention is about a new diagnostic marker for rheumatoid arthritis (RA) that can be used to identify patients with the disease. The marker is anti-MBL autoantibodies, which are found in the serum of RA patients. This new marker is more sensitive and specific than the commonly used rheumatoid factor (RF) and can help detect the level of disease activity and joint damage in RA patients. The invention also provides a diagnostic kit for RA.

Problems solved by technology

The diagnosis of rheumatoid arthritis has been hampered by the lack of a truly disease-specific serologic marker and thus diagnosis of RA is mainly based on clinical criteria recommended by the American College of Rheumatology.
Despite its specificity for RA, because of exacting technical requirements, APF never became widely used.
The tests are done by immunofluorescence but did not become popular in clinical practice, despite high specificity, due to various technical difficulties in performing the assays.

Method used

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  • Novel diagnostic marker, a diagnostic kit and a method for diagnosis of rheumatoid arthritis
  • Novel diagnostic marker, a diagnostic kit and a method for diagnosis of rheumatoid arthritis
  • Novel diagnostic marker, a diagnostic kit and a method for diagnosis of rheumatoid arthritis

Examples

Experimental program
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example 1

[0053] The RA patients were recruited through Army Hospital, Department of Rheumatology, New Delhi, India. The healthy controls were requited mainly through the Guru Teg Bahadur Hospital, Delhi, India. The Army Hospital rheumatologist thoroughly reviewed the medical records of the RA patients and the characterization of the disease was based on the revised criteria of the classification of the disease by American College of Rheumatology Classification criteria. The subject sample included 107 RA affected individuals and 121 normal healthy individual, all from Indian families. The Human Ethics Committee of the Army Hospital, Department of Rheumatology, N. Delhi, Delhi, India and that of the Institute of Genoniics and Integrative Biology, Mall Road, Delhi-110007, India, approved the present study.

example 2

ELISA assay for the Detection of Autoantibodies to MBL

[0054] The presence of anti-MBL autoantibodies was tested for in 107 RA patients and 121 healthy controls. The ELISA tests were conducted with respect to anti-MBL antibodies, using BSA in parallel as the negative control. All the measurements were made in triplicates. The serum of the patient with a higher value of the autoantibodies to MBL was included systematically in each of the assay plates as a positive control.

[0055] The method for the identification of the autoantibodies to MBL in the sera of RA patients and the healthy controls comprises of coating an ELISA plate (Nunc) at 37° C. for two hours with 100 μl / well of serum purified MBL (U.S Biologicals) in a carbonate / bicarbonate-buffer (pH 9.6) at 0.5 μg / ml concentration. Following the steps of washing with Tris-buffered saline (TBS, pH 7.4) containing 0.05% Tween-20 (TBST), blocking of the unoccupied binding sites with 200 μl of 1% bovine serum albumin (BSA) in TBST to...

example 3

ELISA Assay for the Detection of Isotypes of Rheumatoid Factors (IgM RF and IgG RF)

[0060] The presence of isotypes of rheumatoid factors (IgM RF and IgG RF) was measured by ELISA tests in the RA patients and the healthy controls and the standard curve was generated with each assay performed using the serial dilution of the sample used for anti-MBL autoantibody assay.

[0061] ELISA assays were developed for the measurement of rheumatoid factors of IgM and IgG isotypes in the sera of RA patients and the healthy. Flat microtitre plates (Nunc) were coated with 100 μl / well of a 10 μg / ml solution of normal rabbit IgG (Fluka) in carbonate buffer for two hours at 37° C.

[0062] Affecting the binding of rabbit IgG, the unoccupied sites were blocked using 1% BSA for 1 hour at 37° C. Subsequently the plates were washed and incubated with serum samples diluted in TBST for two hours at 37° C. The sample dilution used were 1 / 10, 1 / 50, 1 / 100 and 1 / 1000 for both IgM and IgG RF but the final dilutio...

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Abstract

The present invention relates to a novel diagnostic marker useful for the diagnosis of rheumatoid arthritis comprising the autoantibodies of mannose binding lectin protein and a process thereof.

Description

PRIORITY CLAIM [0001] This application claims priority to Indian Patent Application entitled: “A NOVEL DIAGNOSTIC MARKER, A DIAGNOSTIC KIT AND A METHOD FOR DIAGNOSIS OF RHEUMATOID ARTHRITIS” filed on Nov. 11, 2004. FIELD OF INVENTION [0002] The present invention relates to a novel diagnostic marker useful for the diagnosis of rheumatoid arthritis. [0003] Further, it relates to a method for the diagnosis of rheumatoid arthritis in the human subject. More particularly, it relates to measure the autoantibodies to Mannose binding lectin (MBL) from human serum and quantifying the level of the autoantibodies to the mannose binding lectin (MBL) protein. [0004] The present invention also relates to a diagnostic kit for diagnosis of Rheumatoid arthritis. BACKGROUND OF INVENTION [0005] Rheumatoid arthritis (RA) is a chronic systemic inflammatory disease with peripheral synovitis as its main manifestation. The presentation of the disease and the progression is highly variable both within and b...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N33/537C07K16/42
CPCG01N33/564G01N2333/4724G01N2800/102
Inventor DAS, HASI RANIGUPTA, BHAWNARAGHAV, SUNIL KUMARGOSWAMI, KALYANAGRAWAL, CHARUDAS, RAKHA HARI
Owner COUNCIL OF SCI & IND RES
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