Dendron-mediated DNA virus detection

Inventors

Chae, Chi-Bom • Kim, Kyung-tae • Hur, Seong-Suk

Assignees

Sugentech Inc

Abstract

The disclosure relates to chips containing nucleic acid probes or primers and their use in methods to detect nucleic acid molecules of DNA viruses. The disclosure includes DNA chips with probes immobilized via a dendron-mediated linkage in contact with a thermocycler capable of automatically regulating the temperature, temperature cycle times, and number of temperature cycles of the chips to provide genetic diagnosis in one step.

Core Innovation

The invention relates to a method for detecting a papillomavirus nucleic acid molecule in a biological sample by PCR amplification and in situ hybridization on a solid surface. Amplified nucleic acid material generated by a pair of primers in a PCR reaction mix is denatured in the presence of a single stranded nucleic acid probe complementary to the amplified nucleic acid material and immobilized via attachment to the apex of a conical dendron attached to a solid surface by the base of the conical dendron.

After completing the last cycle of the PCR, the amplified nucleic acid material is denatured within the same PCR reaction mixture to form a denatured mixture. The denatured amplified nucleic acid material is then allowed to hybridize, within the same PCR reaction mixture, to the immobilized probe to form a double stranded complex. The double stranded complex is detected while attached to the solid surface.

A corresponding method is described for detecting a hepadnavirus nucleic acid molecule in a sample with the same dendron-mediated probe immobilization and in situ hybridization workflow. The hepadnavirus nucleic acid molecule is amplified by PCR using a pair of primers in a PCR reaction mix, with a single stranded nucleic acid probe complementary to the amplified nucleic acid material immobilized via a conical dendron on a solid surface, followed by denaturing, hybridizing within the same PCR reaction mixture, and detecting the complex attached to the solid surface.

Claims Coverage

The document provides two independent method claims that share a common dendron-mediated immobilized probe coupled with PCR amplification followed by denaturing and hybridization within the same PCR reaction mixture, followed by detecting the probe-amplicon complex attached to a solid surface. Two inventive features are repeated across the claims, with virus-specific detection for papillomavirus and hepadnavirus.

The claims cover PCR amplification in a reaction mix followed by denaturation and hybridization to an immobilized single stranded nucleic acid probe attached through the apex of a conical dendron on a solid surface, with detection of the resulting double stranded complex while still attached to the solid surface.

Stated Advantages

Documented Applications

No documented applications found