Amplification/detection kit, photothermal PCR amplification method and microorganism detection method using the same

Patent metadata

Field	Value
Publication number	US-12357985-B2
Application number	US-202017100352-A
Country	US
Kind code	B2
Filing date	Nov 20, 2020
Priority date	Sep 18, 2020
Publication date	Jul 15, 2025
Grant date	Jul 15, 2025

Abstract

Disclosed is an amplification/detection kit comprising: a sample pad on which a PCR reaction solution is placed; a transfer pad positioned in contact with the sample pad; a reaction pad into which the PCR reaction solution flowing from the sample pad through the transfer pad flows and which includes a gold thin film deposited on a first surface; and a sealing tape configured to cover and seal a first surface of the transfer pad and a first surface of the reaction pad.

First claim

What is claimed is:

1. A photothermal polymerase chain reaction amplification method using an amplification/detection kit including a gold thin film formed by depositing gold on a pad with an electron-beam, comprising the steps:
surrounding the gold thin film with a sealing frame to withstand an internal pressure of the pad caused by a high temperature during the photothermal polymerase chain reaction amplification method;
a first step of irradiating the gold thin film with light having a predetermined intensity and a predetermined wavelength by using a laser configured to radiate the light having the predetermined intensity and the predetermined wavelength for a first period; and
a second step of irradiating the gold thin film with light having the predetermined intensity and the predetermined wavelength by using the laser for a third period after a second period from an end time point of the first period,
wherein a further step of irradiating the gold thin film with light having the predetermined intensity and the predetermined wavelength by using the laser is not operated for a fourth period from an end time point of the third period,
wherein the gold thin film has a thickness of 10 nm and is deposited on a glass fiber of the pad, and
wherein a photothermal polymerase chain amplification reaction occurs in spaces inside the glass fiber.

2. The photothermal polymerase chain reaction amplification method of claim 1 , wherein:
for the first period and the third period,
heat is transferred to the pad from the gold thin film to isolate DNA of a microorganism from a polymerase chain reaction solution.

3. The photothermal polymerase chain reaction amplification method of claim 2 , wherein:
for the second period and the fourth period,
the heat is emitted from the pad and DNA polymerization and primer annealing of the microorganism occur in the polymerase chain reaction solution.

4. A microorganism detection method using an amplification/detection kit including a gold thin film formed by depositing gold on a pad with an electron-beam, comprising the steps:
surrounding the gold thin film with a sealing frame to withstand an internal pressure of the pad caused by a photothermal polymerase chain reaction amplification process;
a first step of irradiating the gold thin film with light having a predetermined intensity and a predetermined wavelength by using a laser configured to radiate the light having the predetermined intensity and the predetermined wavelength for a first period;
a second step of irradiating the gold thin film with light having the predetermined intensity and the predetermined wavelength by using the laser for a third period after a second period from an end time point of the first period; and
measuring light emitted from the amplification/detection kit after at least the first and second steps of irradiating are terminated,
wherein the gold thin film has a thickness of 10 nm and is deposited on a glass fiber of the pad, and
wherein a photothermal polymerase chain amplification reaction occurs in spaces inside the glass fiber.

5. The microorganism detection method of claim 4 , wherein:
for the first period and the third period,
heat is transferred to the pad from the gold thin film to isolate DNA of a microorganism to be detected from a polymerase chain reaction solution.

6. The microorganism detection method of claim 5 , wherein:
for the second period and the fourth period, the heat is emitted from the pad and DNA polymerization and primer annealing of the microorganism occur in the polymerase chain reaction solution.

Assignees

• Gwangju Inst Science & Tech

Inventors

• Kim Min Gon
• Lee Bo Bin

Classifications

• C12Q1/6851

  Quantitative amplification · CPC title

• B01L2300/0816

  Cards, e.g. flat sample carriers usually with flow in two horizontal directions · CPC title

• B01L2200/10

  Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept · CPC title

• B01L2300/1861

  using radiation · CPC title

• G01N33/569

  for microorganisms, e.g. protozoa, bacteria, viruses · CPC title