Method to increase detection efficiency of real time PCR microarray by quartz material

Patent metadata

Field	Value
Publication number	US-9539571-B2
Application number	US-201013522938-A
Country	US
Kind code	B2
Filing date	Jan 20, 2010
Priority date	Jan 20, 2010
Publication date	Jan 10, 2017
Grant date	Jan 10, 2017

Abstract

A reactor for the quantitative analysis of target nucleic acids using an evanescent wave detection technique and a method for the quantitative analysis of target nucleic acids are provided. The reactor includes a substrate with a cavity, a buffer layer arranged over the substrate, a quartz cover plate arranged over the buffer layer, and inlet and outlet ports. The reactor is thermally and chemically stable for PCR processing and suitable for an evanescent wave detection technique.

First claim

What is claimed is:

1. A reactor for the quantitative analysis of target nucleic acids, comprising:
a substrate having a first planar opposing surface and a second planar opposing surface, the first planar opposing surface of the substrate having a cavity;
a buffer layer comprising a water-impermeable sealant arranged over the first planar surface of the substrate;
a quartz cover plate modified with amino-silane and a functional group, wherein the cover plate is arranged over the buffer layer, the cover plate in combination with the cavity and buffer layer defining a reaction chamber; and
nucleic acid probes tethered to the interior surface of the quartz cover plate in a known, two-dimensional pattern;
wherein the quartz cover plate has less fluorescent background than optical glass k9 for quantitative analysis of target nucleic acids by an evanescent wave detection technique.

2. The reactor of claim 1 , wherein the substrate and the buffer layer are each independently comprised of a chemically inert material that is thermally stable and resistant to contamination.

3. The reactor of claim 1 , wherein the substrate is a glass, a metal, a ceramic, a composite, a polymeric material, or a combination or laminate thereof.

4. The reactor of claim 3 , wherein the polymeric material is a polyimide, polycarbonate, a polyester, a polyamide, a polyether, a polyurethane, a polyfluorocarbon, a polystyrene, a poly(acrylonitrile-butadiene-styrene), a polymethyl methacrylate, polyolefin, or a copolymer thereof.

5. The reactor of claim 3 , wherein the substrate is a thermally conductive polypropylene.

6. The reactor of claim 1 , wherein the substrate has a thermal conductivity greater than about 1 W/mK.

7. The reactor of claim 1 , wherein the water-impermeable sealant is a room temperature vulcanizing silicone rubber.

8. The reactor of claim 1 , further comprising at least one inlet port and at least one outlet port communicating with the reaction chamber through the substrate enabling the passage of fluid from an external source into and through the reaction chamber, and thereby defining a fluid flow path.

9. The reactor of claim 1 , wherein the functional group is a thiocyanate (SCN) functional group.

10. The reactor of claim 1 , wherein the surface of the cover plate includes unreacted SCN groups that are blocked.

11. A reactor for the quantitative analysis of target nucleic acids, comprising:
a substrate having a first planar opposing surface and a second planar opposing surface, the first planar opposing surface of the substrate having a cavity;
a buffer layer arranged over the first planar surface of the substrate;
a quartz cover plate arranged over the buffer layer, the cover plate in combination with the cavity and buffer layer defining a reaction chamber, wherein a surface of the cover plate is modified with amino-silane and further modified with thiocyanate functional groups, and wherein the surface of the cover plate includes unreacted SCN groups that are blocked;
nucleic acid probes tethered to the interior surface of the quartz cover plate in a known, two-dimensional pattern; and
at least one inlet port and at least one outlet port communicating with the reaction chamber through the substrate enabling the passage of fluid from an external source into and through the reaction chamber, and thereby defining a fluid flow path;
wherein the quartz cover plate has less fluorescent background than optical glass k9 for quantitative analysis of target nucleic acids by an evanescent wave detection technique.

12. The reactor of claim 11 , wherein the substrate is a polymeric material and the buffer layer is a water-impermeable sealant, and wherein the substrate has a thermal conductivity greater than about 1 W/mK.

Assignees

• Sun Zhenhong
• Pan Tao
• Liu Xuanbin
• Honeywell Int Inc

Inventors

• Sun Zhenhong
• Pan Tao
• Liu Xuanbin

Classifications

• B01L2300/0816

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

• B01L2300/0636

  Integrated biosensor, microarrays · CPC title

• B01L7/52

  with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples · CPC title

• B01L3/502707

  characterised by the manufacture of the container or its components · CPC title

• B01L3/5027 Primary

  by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip · CPC title