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Method and kit for analyte detection

An analyte and reagent technology, applied in the field of analyte detection

Pending Publication Date: 2018-05-11
GLOBAL LIFE SCI SOLUTIONS OPERATIONS UK LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there are stability issues with the use of short nucleic acids such as specific aptamers and aptamer libraries, and a solution that allows aptamers to remain in the environment before and during the SELEX enrichment, selection, and detection processes is urgently desired. Long-term stability at temperature

Method used

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  • Method and kit for analyte detection
  • Method and kit for analyte detection
  • Method and kit for analyte detection

Examples

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Embodiment 1

[0029] Example 1 - Storage and recovery of small RNAs from chemically coated cellulose-based solid supports

[0030] This example shows that biological samples spotted onto chemically coated cellulose-based solid support matrices can be stored under ambient conditions prior to isolation and purification of single-stranded small molecular weight RNA molecules (~20 nt in length), e.g., by RNA aptamers and microRNAs are exemplified.

[0031] Immediately after whole blood was collected in EDTA by venipuncture, 40 μl aliquots were spotted onto cellulose-based solid supports and allowed to dry. Total RNA was then isolated from the solid support using the Illustra RNAspin MicroRNA Isolation Kit (GE Healthcare) and the Illustra TriplePrep Kit (GE Healthcare). The presence of small molecular weight microRNAs was detected and quantified using two-step quantitative reverse transcription PCR (QRTPCR) with microRNA-specific primers and probes.

[0032] illustra ready-to-use products (GE ...

Embodiment 2

[0062] Example 2 - Extraction and analysis of mRNA molecules from biological samples applied to chemically coated RSM solid supports

[0063] Rat blood collection and dried blood plaque extraction - Male Sprague–Dawley rats (180-250 g) were purchased from Charles River Laboratories. Whole blood was collected via a pipette directly from a 26-gauge catheter placed in the tail vein of anesthetized rats. A 50 microliter aliquot without anticoagulant was transferred and spotted onto chemically treated and untreated filter paper. Blood plaques were dried and maintained at ambient laboratory temperature for 11 days in a desiccator cabinet (~20% relative humidity). Using a 7 mm Harris Uni-Core punch (Fisher Scientific), two sample discs were punched from each dried blood plaque (treated with 15 μl of PK solution (4 mg / ml proteinase K + 0.5% SDS) per punch ). Sample discs were transferred to 1.5 ml microcentrifuge tubes containing 350 μl of extraction solution (RLT buffer with 1%...

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Abstract

The present invention relates to a method and kit for analyte detection. More precisely the invention relates to a method of detecting an analyte, comprising storing short single stranded nucleic acids (10-100 nt), such as aptamers and micro RNA, on a solid support at ambient temperature; and subsequent amplification of said nucleic acids for detection of analyte(s).

Description

field of invention [0001] The present invention relates to methods and kits for analyte detection. More precisely, the present invention relates to methods for detecting analytes comprising storing short single-stranded nucleic acids (10-100 nt), such as aptamers and microRNAs, on solid supports at ambient temperature; and subsequently amplifying them with The nucleic acid used to detect the analyte. Background of the invention [0002] Biological sample storage and preservation is desirable as preserved samples can be used in various applications such as analyte detection, sensing, forensic and diagnostic applications, genome sequencing, whole genome amplification, etc. [0003] Several devices are currently available for storing samples on paper substrates. Porous or non-porous substrates are often used to preserve biological samples, such as paper cards or membranes. Examples of paper cards or membranes include chemically treated FTA for preservation of nucleic acid sa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/686
CPCC12Q1/6834C12Q2525/204C12Q2531/113C12Q2525/205C12Q2525/207C12N15/1048C12N15/115C12N2310/16C12N2320/13C12N2330/31
Inventor J.K.霍尔顿P.J.塔特内尔
Owner GLOBAL LIFE SCI SOLUTIONS OPERATIONS UK LTD
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