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Methods and devices relating to the detection of oral cancer biomarkers

A biomarker and oral cancer technology, applied in biochemical cleaning devices, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problem of not providing oral sample device OSCC

Active Publication Date: 2017-08-18
GLOBAL LIFE SCI SOLUTIONS OPERATIONS UK LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are currently no oral sample devices or assays commercially available to detect or predict OSCC progression

Method used

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  • Methods and devices relating to the detection of oral cancer biomarkers
  • Methods and devices relating to the detection of oral cancer biomarkers
  • Methods and devices relating to the detection of oral cancer biomarkers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Direct measurement of interleukin (IL) from a solid support

[0037] Recombinant IL-2±vector (R & D Systems; Cat. 202-IL-CF-10 μg; lot AE4309112 and Cat. 202-IL-10 μg; lot AE4309081, respectively) was dissolved in blood at 50 pg or 100 pg / μl (TCS Biosciences). Aliquots (1 μl containing 50 (B) or 100 (A) pg IL-2) were applied to Whatman 903 filter paper.

[0038] The samples were allowed to dry overnight at ambient temperature and humidity. Using an appropriately sized punch, a 3 mm diameter punched disk was extracted from each paper type. Individual discs were directly assayed for IL-2 using reagents from the complete 10-configuration IL-2 QuantikineELISA kit (R & D Systems, Cat. D2050, lot 273275). Direct assays are performed "punch in well", ie, where sections 903 of filter paper are punched out and deposited in reaction wells of conventional multiwell plates.

[0039] Upon completion of the assay, the optical density was monitored at 450 nm. IL-2 reco...

Embodiment 2

[0040] Example 2: Model enzyme assays from cells or enzymes transferred to solid supports

[0041] Protein and enzyme assays were performed using fully configured DNase and RNase Contamination kits (DNase & RNAase Alert QC Systems, cat# AM1970 & AM1966, Life Technologies) according to the manufacturer's instructions.

[0042] Dideoxyribonuclease (DNase)

[0043] In the first series of experiments, 0.125-0.5 U DNase was applied to Whatman FTA and 903 paper in a volume of 10 µl. DNase and RNase activities were measured as outlined below.

[0044] In a second series of experiments, 1.2 mm punches were taken from 106 human embryonic stem cells (GE Healthcare; cell line ref: WCB307 GEHC 28) that had been applied to FTA and 903 paper in a volume of 10 μl as above. DNase and RNase activities were measured as outlined below.

[0045] In a third series of experiments, from 106 human embryonic stem cells (GEHealthcare; cell line ref: WCB307 GEHC 28) that had been applied to FTA and...

Embodiment 3

[0049] Example 3: Measuring Lactate Dehydrogenase (LDH)

[0050] Enzyme assay for LDH

[0051] LDH is a cytosolic enzyme present in many different types of cells. Upon injury to the plasma membrane, LDH is released into the bathing medium surrounding the cell. Released LDH can be quantified by coupled enzyme reactions. First, LDH catalyzes the conversion of lactate to pyruvate by reducing NAD+ to NADH. Second, diaphorase uses NADH to reduce tetrazolium salt (INT) to red formazan product. Therefore, A The level produced is directly proportional to the amount of LDH released in the medium. Assays were performed by transferring the punch and adding cells or LDH enzyme or into a microplate and adding kit reagents (LDH Cytotoxicity Assay Kit; Thermo Scientific, Product Code 88953). After incubation for 30 minutes at room temperature, the reaction was stopped and LDH activity was determined by spectrophotometric absorbance at 490 nm.

[0052] Immunoassay for LDH

[00...

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Abstract

Disclosed is an ex vivo method for detecting plural predefined oral biomarkers within biological material present in a human oral cavity when oral cancerous activity is present in said cavity, the method comprising the steps of: i) providing a solid support for accepting a sample of biological material from the oral cavity; ii) transferring a sample of the biological material to the solid support, iii) performing one or more ex vivo plural assays to detect the presence of said plural biomarkers on the solid support, in the form of one or more nucleic acid sequences, and / or one or more proteins, and / or one or more enzymes indicative of said oral cancerous activity. A sample swab 10 suitable for use with the above methods is disclosed also.

Description

[0001] field of invention [0002] The present invention relates to an ex vivo method for the detection and quantification of various oral biomarkers in biological material present in the human oral cavity in the presence of oral cancer activity. [0003] Background of the invention [0004] Oral cancer has become a public health concern with increased morbidity and mortality worldwide. Oral cancer (primarily oral squamous cell carcinoma, OSCC) is the sixth most common malignancy in humans, with a high incidence and a five-year mortality rate approaching 50%, which has not changed significantly in more than 50 years. Therefore, it is extremely important to implement newer screening and early detection methods, which can reduce the morbidity and mortality associated with this disease. The current treatment modalities offered to OSCC patients are based on tumor metastasis criteria and histological grade. Unfortunately, these predictors are subjective and relatively unreliable b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/34G01N33/566
CPCG01N33/57407A61B5/150053C12P19/34C12Q1/6806C12Q1/6886C12Q2600/158G01N33/574G01N2800/7028
Inventor J.K.霍尔顿P.J.塔特内里
Owner GLOBAL LIFE SCI SOLUTIONS OPERATIONS UK LTD
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