Patent Application
20080199862
Embodiments of the present invention relate to an active biochip for nucleic acid preparation. More specifically, embodiments of the invention relate to an active biochip for DNA analysis utilizing a hydroxyapatite chromatographic adsorption column.
Nucleic acid diagnosis is an important and blooming part of in vitro diagnosis (IVD), in which polymerase chain reaction (PCR) and other nucleic acid amplification techniques are the principal diagnosis tool. Nucleic acid (e.g., DNA) microarrays are currently being explored as an interesting technology platform for future IVD products. However, there are still technological bottlenecks to be resolved in the nucleic acid testing process. These problems hinder the goal for making the analysis more simple, robust, rapid and reproducible. Sample handling represents one of the main bottlenecks. It usually takes more than one man-hour to prepare a dozen nucleic acid samples from clinical samples. Another challenge to a medical lab is to process the thousands of samples waiting for nucleic acid preparation on a daily basis. More importantly, heavy sample preparation work will often induce cross-contamination and false positive or false negative results. This causes a low reproducibility and high variation in the testing.
In the drawings, which are not necessarily drawn to scale, like numerals describe substantially similar components throughout the several views. Like numerals having different letter suffixes represent different instances of substantially similar components. The drawings illustrate generally, by way of example, but not by way of limitation, various embodiments discussed in the present document.
Embodiments of the invention relate to an active biochip for nucleic acid analysis. The biochip comprises an inlet for introducing a nucleic acid sample, fluid channels, valves in contact with the fluid channels and pumps in contact with the fluid channels and adapted to generate a carrier gas or move a buffer through a portion of the fluid channels. The biochip also includes one or more hydroxyapatite columns for separating a portion of the nucleic acid sample, buffer reservoirs in contact with the fluid channels and positioned near the pumps, air exits, a waste reservoir and a nucleic acid analysis region.
Embodiments of the present invention also relate to a method of analyzing a nucleic acid sample utilizing an active biochip. The method comprises introducing a nucleic acid sample to a fluid channel in the biochip, activating a carrier gas sufficient to move the nucleic acid sample through the fluid channel, binding at least a portion of the sample on a hydroxyapatite column, removing unspecific binding substances from the column, releasing the bound portion of the sample sufficient to provide a released sample and analyzing the released sample.
The following detailed description includes references to the accompanying drawings, which form a part of the detailed description. The drawings show, by way of illustration, specific embodiments in which the invention may be practiced. These embodiments, which are also referred to herein as “examples,” are described in enough detail to enable those skilled in the art to practice the invention. The embodiments may be combined, other embodiments may be utilized, or structural, and logical changes may be made without departing from the scope of the present invention. The following detailed description is, therefore, not to be taken in a limiting sense, and the scope of the present invention is defined by the appended claims and their equivalents.
In this document, the terms “a” or “an” are used to include one or more than one and the term “or” is used to refer to a nonexclusive “or” unless otherwise indicated. In addition, it is to be understood that the phraseology or terminology employed herein, and not otherwise defined, is for the purpose of description only and not of limitation. Furthermore, all publications, patents, and patent documents referred to in this document are incorporated by reference herein in their entirety, as though individually incorporated by reference. In the event of inconsistent usages between this document and those documents so incorporated by reference, the usage in the incorporated reference should be considered supplementary to that of this document; for irreconcilable inconsistencies, the usage in this document controls.
Embodiments of the invention relate to an active biochip for nucleic acid preparation. The biochip allows for a simple, robust, rapid and reproducible nucleic acid testing system. The biochip utilizes hydroxyapatite (HA) chromatographic absorption for nucleic acid preparation that is capable of preparing pure nucleic acid in an automated fashion for PCR and microarray analysis. Because hydroxyapatite is a biocompatible material capable of specific binding with nucleic acid in a high salt condition, it can be integrated into a microfluidic biochip for purification of nucleic acid. The active microfluidic biochip is laborsaving and whole-sealed, which reduces the operative error and cross-contamination and increases the reliability of the analytical results. The nucleic acid sample preparation/analysis may be automated and completed in around 10 minutes, rather than hours.
Referring to
First valve 102 and seventh valve 104 may be in an open position when introducing a sample that contains a nucleic acid into the inlet 106. Valves 102 and 104 may then be closed and the fifth valve 108 may be opened. Pump 110 may then be activated, such as by heating to generate a carrier gas. The sample may then be pushed through the HA column 130 and into the waste reservoir 132. The target nucleic acid will selectively bind to the column 130. Pump 110 may then be stopped and the third pump 122 activated (valve 118 now open). Pump 122 will then push a high salt buffer (from reservoir 120) through the column 130 to wash any unspecific binding substance from the column to the waste reservoir 130. Pump 122 may then be stopped and then the washing may be repeated with pump 134, while opening valve 128. Valves 118, 128 and 108 may be closed and valves 112 and 136 opened. Pump 116 may then be activated to push a low salt buffer from reservoir 114 through the column 130, which releases the bound target nucleic acids in the sample. The released sample then flows to the nucleic acid analysis region 136. The valves, heaters, pumps and analysis region may all be controlled by onboard chip circuitry.
Referring to
The solid chemical propellant 212 may include azobis-isobutyronitrile (AIBN), for example. The propellant 212 may be in powder form. The solid chemical propellant 212 may be heated to produce a gas, such as nitrogen. The output pressure of the gas, generated from the solid chemical propellant, may be adjustable to a desired pressure by controlling the input power of the heater. The gas may be utilized as a carrier gas to push a fluid sample through the fluid channel.
The layers 202, 204, 206 and 208 may be manufactured of an inexpensive plastic, such as polymethyl methacrylate (PMMA) or polydimethylsiloxane (PDMS), for example. The second 204 and third layers 206 may be crosslinked in certain areas, to support porous membranes or channel structure, for example. The four layers may be bonded by plastic hot embossing processes.
Referring to
A hydrogel is a network of hydrophilic polymers that can swell in water and hold a large amount of water while maintaining their structure. A three-dimensional network is formed by cross-linking polymer chains. The hydrogel 406 utilized as a valve may be temperature sensitive. Examples of such hydrogels include poly(N-iso-propylacrylamide) (PNIPAAm) and poly(N,N-diethylacrylamide) (PDEAAm), for example.
Referring to
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The sample may be lysed tissue, for example. The sample may be introduced 602 by injection. The carrier gas may be activated 604 by heating a solid chemical propellant to generate the carrier gas. The unspecific binding substances may be removed 608 from the column by contacting with a high salt buffer, such as by washing. Releasing 610 the bound sample may include contacting the sample with a low salt buffer. The nucleic acid analysis region may include a microarray or a PCR region in conjunction with a microarray, for example. The sample may be analyzed 612 by amplification or by microarray detection methods.
The Abstract is provided to comply with 37 C.F.R. §1.72(b) to allow the reader to quickly ascertain the nature and gist of the technical disclosure. The Abstract is submitted with the understanding that it will not be used to interpret or limit the scope or meaning of the claims.
