Portable micro-fluidic chip LAMP (loop-mediated isothermal amplification) visible detector and detection method thereof

A technology of microfluidic chips and detectors, applied in chemical instruments and methods, biochemical equipment and methods, biochemical instruments, etc., can solve the problems of not reaching POCT, and achieve the effect of saving time

Inactive Publication Date: 2017-03-22
RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the above research still needs auxiliary equipment such as water baths and hot plates to provide constant temperature, which cannot meet the requirements of POCT due to the limitation of fixed testing places.

Method used

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  • Portable micro-fluidic chip LAMP (loop-mediated isothermal amplification) visible detector and detection method thereof
  • Portable micro-fluidic chip LAMP (loop-mediated isothermal amplification) visible detector and detection method thereof
  • Portable micro-fluidic chip LAMP (loop-mediated isothermal amplification) visible detector and detection method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Step 1. Making a microfluidic chip 5

[0057] The SU-8 positive mold is processed by soft lithography technology, and the microfluidic chip 5 is produced by molding method. PDMS is poured on the positive mold, and bonded with the glass sheet after curing.

[0058] Among them, the LAMP reaction area of ​​the chip 5 has five detection channels 21-25 and corresponding independent control channels P1-P5, each channel has a volume of about 7 µL, and its size is 10 mm (L) × 2.4 mm ( W) × 0.3 mm (H). Each detection channel 21-25 is connected to the meandering microchannel of the fluid control channel 30, and its size is 29.5 mm (L) × 0.1-0.2 mm (W) × 0.05-0.3 mm (H); a positive control channel P is set at the same time , negative control channel N and negative independent control channel N*, as the quality control of the reaction system. Liquid mineral oil is used for fluid control during the use of the microfluidic chip 5 .

[0059] Step 2. Process the portable microfluidi...

Embodiment 2

[0083] The application of the microfluidic chip 5 includes detection of multi-site gene mutations in myeloproliferative tumors, multi-site gene mutations in other tumors, and pathogen nucleic acid detection.

Embodiment 3

[0085] The application of the visualized detector covers all application fields involved in the isothermal amplification of microfluidic chips, including the detection of transgenic product safety, virus detection, common pathogenic bacteria detection, bacterial drug resistance gene detection and tumor detection .

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Abstract

The invention relates to a portable micro-fluidic chip LAMP (loop-mediated isothermal amplification) visible detector and a detection method thereof. The visible detector comprises a box cover and a box body, wherein the box cover and the box body are connected by virtue of a locking plate on the side face; a micro-fluidic chip, a temperature control device and a visible detection system are arranged in the box body; the temperature control device comprises a temperature controller and a heating metal block; the micro-fluidic chip is arranged on the heating metal block; a transparent stopper is arranged around the micro-fluidic chip and the heating metal block; a detachable cover plate is arranged on the transparent stopper; the visible detection system is arranged on the cover plate; and the visible detection system comprises a micro camera and a micro ultraviolet fluorescent lamp. The detection method of the visible detector comprises steps of manufacturing the micro-fluidic chip, injecting a primer and reaction fluid, conducting an isothermal reaction and reading a detection result. The invention further relates to an application of the visible detector. The technical scheme provided by the invention, in comparison with the prior art, achieves the instant detection of an LAMP reaction, so that user's time is effectively saved.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to a portable microfluidic chip LAMP visual detector and a detection method thereof. Background technique [0002] Loop-mediated isothermal amplification (LAMP) is a new technique for in vitro isothermal amplification of specific nucleic acid fragments. This technology mainly uses two pairs of specially designed primers and a DNA polymerase with strand displacement activity to make circular single-strand structures appear in the reaction at the primer junctions at both ends of the template, so as to ensure that the primers can be smoothly combined with the template under isothermal conditions. And carry out strand displacement amplification reaction. Because it overcomes the shortcomings of the traditional polymerase chain reaction (PCR reaction) to obtain single-stranded templates through repeated thermal denaturation processes, and avoids the time-consuming process of repeated cooli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/00C12M1/38C12M1/34C12Q1/68
CPCB01L3/5027B01L2300/1805C12Q1/6844C12Q2531/119C12Q2563/107C12Q2565/629
Inventor 李敏汪骅刘倩徐燕萍马晓惟邬叶红
Owner RENJI HOSPITAL AFFILIATED TO SHANGHAI JIAO TONG UNIV SCHOOL OF MEDICINE
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