System for integrated analysis of real-time polymerase chain reaction and DNA chip and method for integrated analysis using the same
Inventors
Seo, Sung-min • Lee, Do-Bu • Lee, Joong Hwan • Paek, Mun-Cheol • Ku, Su-Jin
Assignees
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Publication Number
US-9551038-B2
Publication Date
2017-01-24
Expiration Date
Abstract
Provided are a system for integrated analysis of a real-time polymerase chain reaction and a DNA chip and a method for integrated analysis using the same, and more particularly to an apparatus for integrated analysis of a real-time polymerase chain reaction and a DNA chip and a method for integrated analysis using the same. According to the method for integrated analysis of a biomaterial of the present invention, gene amplification proceeds and subsequently hybridization proceeds in a single reactor, thereby preventing contamination of the sample due to external factors, which may be caused while the sample is transferred for reaction, and automating a series of procedures such as injection of the sample, reaction of the biomaterial, and detection and analysis of results.
Core Innovation
The invention provides a method for integrated analysis of a biomaterial in a sample using a biomaterial detecting device, characterized by simultaneously performing quantitative analysis of a reaction product amplified by real-time PCR and qualitative analysis of the reaction product by a biochip having a target gene probe immobilized thereon in a single reactor. The method performs gene amplification in the single reactor and conducts hybridization of the amplification reaction product with the target gene probe in the single reactor only through temperature change, enabling integrated analysis in one reactor for both quantitative and qualitative results.
The biomaterial detecting device includes a reactor, a head, and a reactor coupler protruded downwardly from the head. The reactor coupler has a first protrusion and a second protrusion formed on a first portion and a second portion of an outer circumferential surface of the reactor coupler, respectively, and protruded from the bottom surface of the head. The protrusions are formed so that the protrusion prevents the head from contacting the reactor and prevents all parts of the reactor coupler inserted into the inside of the reactor, with a space between the first protrusion and the second protrusion.
The device also includes a rod extended downwardly from the reactor coupler, and the biochip is formed on the lower side of the rod. The reactor coupler is coupled with a reactor such that a part of the reactor coupler is inserted into an inside of the reactor and the rest of the reactor coupler is not inserted into the inside of the reactor. The rod is detachable from the reactor coupler to allow replacement of the rod.
Claims Coverage
The claims include one independent method claim with device structural limitations, covering integrated quantitative real-time PCR and qualitative biochip hybridization performed in a single reactor with temperature change only, together with a detachable rod and a specific reactor coupler interface structure using protrusions to control contact.
Integrated single-reactor quantitative real-time PCR and qualitative biochip hybridization
Simultaneously performing quantitative analysis of a reaction product amplified by using a real-time PCR and qualitative analysis of the reaction product by a biochip having a target gene probe immobilized thereon in a single reactor, comprising injecting a real-time PCR reagent and a target gene primer into the single reactor for gene amplification, and conducting hybridization of the amplification reaction product with the target gene probe in the single reactor only through the temperature change.
Reactor coupler with protrusions preventing contact during coupling
Providing a biomaterial detecting device comprising a reactor, a head, and a reactor coupler protruded downwardly from the head, where the reactor coupler has a first protrusion and a second protrusion on outer circumferential surface portions and protrudes from the bottom surface of the head, with a space between the first protrusion and the second protrusion, and the protrusions are formed so that the protrusion prevents the head from contacting the reactor and prevents all parts of the reactor coupler inserted into the inside of the reactor.
Detachable rod carrying the biochip
Forming the device to include a rod extended downwardly from the reactor coupler, with the biochip formed on the lower side of the rod, where the rod is detachable from the reactor coupler to allow replacement of the rod.
Partial insertion coupling of reactor coupler
Coupling the reactor coupler with a reactor so that a part of the reactor coupler is inserted into an inside of the reactor and the rest of the reactor coupler is not inserted into the inside of the reactor.
Overall, claim coverage centers on a single-reactor workflow that combines quantitative real-time PCR with qualitative biochip hybridization using temperature change only, together with a biomaterial detecting device architecture that includes a reactor coupler with protrusions to avoid contact during partial insertion coupling and a detachable rod carrying the biochip.
Stated Advantages
Documented Applications
Detecting Mycobacterium tuberculosis presence/antibiotic resistance and drug-resistant hepatitis B virus using the integrated method and device.
Detecting antibiotic resistance related to rifampin (rpoB) and isoniazid (inhA/katG) for Mycobacterium tuberculosis.
Detecting lamivudine resistance associated with drug-resistant hepatitis B virus via HBV DNA polymerase mutations in the YMDD region (as described with codons 528, 529, 514, 552, 548, 555).
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