Genetec Inc.

Marilyn J Stapleton

Warren R Jewett

Rebecca Sundseth

Villette S Thorpe

William Harrington

Ke Wei

The invention is a method and device for collecting and processing a biological specimen for the analyses of nucleic acids. A device comprises a matrix to which cells and viruses adhere and a handle to manipulate the matrix. The devices are used to collect, dry, transport, store and process small amounts of blood or other tissue. The matrix of the device is transferred to a reaction tube and amplifying reagents added to it. Nucleic acid sequences and relative quantities are detected and analyzed from the same specimen. The relative amounts of amplified nucleic acid from one or more particular RNA sequences are compared to one another and to the amount of amplified nucleic acid from DNA sequences serving as an internal control for the number of biological units per specimen. The relative amounts of amplified viral sequences from suspected viruses in the biological specimen and from recombinant viral particles serving as a viral quantitation standard enable estimation of viral burden in a given quantity of specimen.

Immobilizing and processing specimens on matrix materials for the

The invention is an apparatus and a process for containing and handling liquids in a thin capillary space. Two opposing walls positioned at a first and a second distance apart define a thin space having at least two different heights and two different regions of capillary liquid attraction. Enclosing the thin space with side walls defines a reaction chamber. A liquid receiving area, entry and exit ports and features projecting from, or encircling, at least one wall allow coatings on the internal surface of either of the two opposing walls to react with multiple microvolumes of reagents and wash treatments in series. Liquids flow into, or are withdrawn from, the thin space in a way in which bubbles are not introduced into the thin space. Bubbles that may arise in the thin space move along a sloped feature out of the thin space into an annular enclosure. The liquids are withdrawn after reacting with the coatings, either to empty the chamber and recover the liquid volume, or to wash the coatings with treatment liquids and introduce new treatment liquids. The ability to control liquid movement in spaces thin enough to exhibit capillarity is useful for analyzing biological specimens cost-effectively with less reagent volume and more information per specimen and without sacrificing convenience in liquid handling or surface reaction space.

Thin reaction chambers for containing and handling liquid microvolumes

Device for processing biological specimens for analysis of nucleic acids

A process for handling a biological specimen for the analysis of nucleic acid sequences wherein the biological specimen is immobilized within a carrier device for controlled temperature conditions and the sequential addition of fluid treatments. The fluid treatments arm selected from lysing end denaturing solutions, wash or rinse solutions, reagents for nucleic acid amplification, electrophoresis and hybridization and labeling and detection reagents. The fluid treatments are unique to each specimen and the detection of target nucleic acid sequences is localized within the carrier device.

Process for specimen handling for analysis of nucleic acids

An apparatus for the automated detection of target nucleic acid sequences in which multiple biological samples are individually incorporated into a matrix and treated in a stepwise fashion to expose, amplify, detect and enumerate the presence of original, target nucleotide sequences in each sample. The samples are held together with other samples in a chamber connected by means of a fluid flow conduit to which the matrices make connection via numerous jet-spray ports for the injection of various fluids from a plurality of reservoirs used to process the samples. Solvents and reagents are used to degrade and wash cellular structures and contaminants away from the nucleic acids. Primer and polymerase molecules are used to replicate the target sequences of nucleic acids in the sample. Labeled DNA probe molecules serve to hybridize to the amplified target sequences for detection. Appropriate buffers allow these manipulations on the nucleic acid molecules in the samples. The system also includes a pump to draw the fluids from the reservoirs and force them through the jet spray manifolds into the main chamber and valves under the control of a microprocessor. The system also includes fans and heating elements for regulating temperature and humidity. Different nucleic acid targets in different biological specimens are accommodated by regulating the quantity, duration, pH, temperature and pressure of fluids flowing from the reservoirs.

Method for studying nucleic acids within immobilized specimens

A cover has been designed to make a sealed chamber over a desired specimen area to control reagent evaporation which is critical for amplifying DNA in slide specimens. The cover is positioned over the specimen and temporarily adhered to, or pushed against, the slide, forming a sealed chamber. Reagents are added through a channel in the cover with a standard pipette and the channel opening is closed to prevent evaporative loss of reagents. The chamber is also designed to relieve internal vapor pressure, which is creating upon heating, thereby facilitating diffusion in and out of cells rather than allowing high pressure to flatten cells against the slide surface. Increasing the height of the chamber upwards toward a neck in the cover allows air bubbles being formed during each cycle to leave the specimen area, while at the same time preventing water vapor loss or partitioning of water vapor condensate.

Devices for containing biological specimens for thermal processing

An apparatus and method for performing automated sample preparation, DNA amplification and detection, which apparatus has heat-sinking, flat carriers for holding specimens and reagents, devices for heating and cooling and maintaining the specimen to or at any given temperature for any given time periods, and a computer to generate signals that control said temperatures and times.

Temperature control apparatus and method

The matrix carrier is a hinged compartment facilitating automation of DNA- and RNA-based diagnostics and genetic surveillance and detection. Specimens are embedded in a matrix in the carrier. The matrix is then treated by one or more of the techniques such as amplification, electrophoresis, and hybridization as selected for the desired analysis and then the sample is treated to detect the cellular component.

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Durham, NC, United States of America

Marilyn J Stapleton