Rapid antibiotic susceptibility testing system based on bacterial immobilization using gelling agent, antibiotic diffusion and tracking of single bacterial cells
원문보기
IPC분류정보
국가/구분
United States(US) Patent
등록
국제특허분류(IPC7판)
C12Q-001/18
C12N-011/04
B29C-045/00
B29L-031/00
출원번호
US-0793740
(2015-07-08)
등록번호
US-9920351
(2018-03-20)
우선권정보
KR-10-2012-0011002 (2012-02-02)
발명자
/ 주소
Kwon, Sunghoon
Jung, Yong-Gyun
Choi, Jung Il
Na, Hun Jong
출원인 / 주소
QUANTAMATRIX INC.
대리인 / 주소
STIP Law Group, LLC
인용정보
피인용 횟수 :
1인용 특허 :
6
초록▼
A testing method is disclosed. The testing method includes: providing a mixture solution of a gelling agent and a microbe to a gelling device; solidifying the mixture solution to form a solid thin film in which the microbe is immobilized; supplying a bioactive agent to the solid thin film and allowi
A testing method is disclosed. The testing method includes: providing a mixture solution of a gelling agent and a microbe to a gelling device; solidifying the mixture solution to form a solid thin film in which the microbe is immobilized; supplying a bioactive agent to the solid thin film and allowing the bioactive agent to diffuse into the solid thin film; and imaging the individual responses of the single microbial cells to the bioactive agent, and determining the minimum inhibitory concentration (MIC) of the bioactive agent based on the analysis of the images to obtain AST results.
대표청구항▼
1. A method for observing the responses of one or more microbial cells to a bioactive agent, comprising: providing a mixture solution of a gelling agent-containing liquid medium and one or more microbial cells;gelling the mixture solution to form a gelled matrix wherein the one or more microbial cel
1. A method for observing the responses of one or more microbial cells to a bioactive agent, comprising: providing a mixture solution of a gelling agent-containing liquid medium and one or more microbial cells;gelling the mixture solution to form a gelled matrix wherein the one or more microbial cells is immobilized inside the gelled matrix;providing a bioactive agent into the gelled matrix and allowing the bioactive agent to diffuse into the gelled matrix so that the bioactive agent is delivered to the one or more microbial cells;tracking at least one individual microbial cell directly among the one or more microbial cells by a microscopy and imaging an increase in an area occupied by growth of the at least one individual microbial cell with the lapse of time; andanalyzing the time-lapse images. 2. The method according to claim 1, wherein a thickness of the gelled matrix is adjusted and/or optical focusing is performed for imaging the single microbial cells are observed. 3. The method according to claim 1, wherein the imaging is carried out to determine growth, division or aggregation of the one or more microbial cells. 4. The method according to claim 1, wherein the gelling agent is selected from the group consisting of agar, agarose, gelatin, alginate, collagen, fibrin, and mixtures thereof. 5. The method according to claim 1, wherein the gelled matrix is formed using a gelling device that has at least one receiving portion having an internal space where the mixture solution is retained. 6. The method according to claim 5, wherein the receiving portion comprises a microfluidic channel structure. 7. The method according to claim 6, wherein the microfluidic channel is hydrophilized. 8. The method according to claim 1, wherein the imaging step further comprises observing changes in the growth of the one or more microbial cells depending on a concentration of the diffused bioactive agent and the analyzing is carried out on the obtained images. 9. The method according to claim 8, wherein the imaging is carried out to observe a contact interface between the one or more microbial cells-immobilized in the gelled matrix and the bioactive agent, and the analysis is carried out on the obtained images. 10. The method according to claim 8, wherein a minimum inhibitory concentration (MIC) or a minimum biofilm eradication concentration (MBEC) of the bioactive agent is determined based on analysis of the images. 11. The method according to claim 8, wherein the bioactive agent susceptibility and resistance of the one or more microbial cells are determined based on analysis of the images. 12. The method according to claim 1, wherein the responses of the one or more microbial cells are analyzed by at least one of a number of one or more microbial cells, occupied area, and cell growth morphology of the one or more microbial cells in the images. 13. A method for observing the responses of one or more microbial cells to a bioactive agent, comprising: providing a bioactive agent into a microbe-immobilized solid thin film based on at least one gelling agent selected from the group consisting of agar, agarose, gelatin, alginate, collagen, and fibrin and allowing the bioactive agent to penetrate into the solid thin film by diffusion so that the bioactive agent is delivered to the one or more microbial cells; andtracking at least one individual microbial cell directly among the one or more microbial cells with an optical measurement system and imaging an increase in an area occupied by growth of the at least one individual microbial cell. 14. The method according to claim 13, wherein the solid thin film has a thickness of 1 μm to 5 mm. 15. The method according to claim 13, wherein the optical measurement system comprises an imaging system adapted to obtain images from targets and an image processing unit adapted to process and analyze the obtained images. 16. The method according to claim 13, wherein the changes in the growth of the one or more microbial cells are analyzed by calculating areas occupied by the one or more microbial cells in the images. 17. A method for observing the responses of one or more microbial cells to a bioactive agent, comprising: contacting a gelling agent-containing liquid medium with the one or more microbial cells;solidifying the gelling agent-containing liquid medium to form a gelled matrix in which the one or more microbial cells are immobilized by the gelled matrix;providing a bioactive agent into the gelled matrix and allowing the bioactive agent to diffuse into the gelled matrix so that the bioactive agent is delivered to the immobilized one or more microbial cells;incubating the immobilized one or more microbial cells for a certain time; andtracking at least one individual microbial cell directly among the one or more microbial cells by a microscopy and imaging an increase in an area occupied by growth of the at least one individual microbial cell. 18. The method according to claim 17, wherein the imaging step further comprises observing changes in growth of the microbe depending on a concentration of the diffused bioactive agent. 19. The method according to claim 17, wherein the responses of the microbe are analyzed by at least one of number, occupied area, and cell growth morphology of the one or more microbial cells in the images. 20. A method for an antibiotic susceptibility testing (AST), comprising: providing one or more microbial cells;providing a gelling agent capable of immobilizing the one or more microbial cells, wherein the gelling agent is selected from the group consisting of agar, agarose, gelatin, alginate, collagen, fibrin, and mixtures thereof;providing a bioactive agent capable of inhibiting growth of the one or more microbial cells;forming a solid thin film comprising the one or more microbial cells, the gelling agent, and the bioactive agent, wherein the one or more microbial cells are immobilized by the gelling agent;incubating the one or more microbial cells;tracking at least one individual microbial cell directly among the one or more microbial cells by a microscopy and imaging an increase in an area occupied by growth of the at least one individual microbial cell according to an incubation time and a concentration of the bioactive agent, wherein an image from the imaging step is taken as a microscopic picture from the microscopy;based on the image, determining a minimum inhibitory concentration (MIC) by analyzing the occupied area or a morphology based on the growth of the at least one individual microbial cell; andobtaining the result of the AST against the bioactive agent based on the step of determining the MIC. 21. A method for analyzing one or more microbial cells interacting with a bioactive agent, comprising: providing one or more microbial cells;providing a gelling agent capable of immobilizing the one or more microbial cells, wherein the gelling agent is selected from the group consisting of agar, agarose, gelatin, alginate, collagen, fibrin, and mixtures thereof;providing a bioactive agent capable of inhibiting growth of the one or more microbial cells;forming a solid thin film comprising the one or more microbial cells, the gelling agent, and the bioactive agent, wherein the one or more microbial cells are immobilized by the gelling agent;incubating the one or more microbial cells; andtracking at least one individual microbial cell directly among the one or more microbial cells by a microscopy and imaging an increase in an area occupied by growth of the at least one individual microbial cell.
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이 특허에 인용된 특허 (6)
Kwon, Sunghoon; Jung, Yong-Gyun; Choi, Jung Il; Na, Hun Jong, Rapid antibiotic susceptibility testing system based on bacterial immobilization using gelling agent, antibiotic diffusion and tracking of single bacterial cells.
Goldberg, David A.; Howson, David C.; Metzger, Steven W.; Buttry, Daniel A.; Saavedra, Steven Scott, Sensitive and rapid determination of antimicrobial susceptibility.
Goldberg, David A.; Howson, David C.; Metzger, Steven W.; Buttry, Daniel A.; Saavedra, Steven Scott, Sensitive and rapid determination of antimicrobial susceptibility.
Goldberg, David A.; Howson, David C.; Metzger, Steven W.; Buttry, Daniel A.; Saavedra, Steven Scott, Sensitive and rapid determination of antimicrobial susceptibility.
Paul M. Matsumura ; Jones M. Hyman ; Scott R. Jeffrey ; Martin J. Maresch ; Thurman C. Thorpe ; William G. Barron, Susceptibility plates for microbial antibiotic susceptibility testing.
Kwon, Sunghoon; Jung, Yong-Gyun; Choi, Jung Il; Na, Hun Jong, Rapid antibiotic susceptibility testing system based on bacterial immobilization using gelling agent, antibiotic diffusion and tracking of single bacterial cells.
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