Abstract: Described are methods, kits and systems for multiplexed detection of biological agents in a sample, e.g., multiplexed detection of e.g., bacteria, viruses, and biological toxins. The method utilizes two markers for each agent; the presence or absence in a sample of each of the two markers per agent is determined in separate reactions; however each reaction is used to detect a single marker for multiple agents. Also disclosed is the multiplexed detection method using real time PCR. The invention provides an efficient, cost-effective, and specific method for multiplexed detection of biological agents.

Abstract: The present invention provides methods, apparatuses and computer programs for verifying the integrity of a probe by comparing the fluorescence value of a probe to a threshold value. The invention also provides for methods, apparatuses and computer programs for normalizing the fluorescence value of a probe and detecting a target nucleic acid in a sample.

Abstract: This invention provides an apparatus for rapidly heating and/or cooling a sample in a reaction vessel. In some embodiments, the apparatus includes optics for the efficient detection of a reaction product in the vessel. The invention also provides a reaction vessel having a reaction chamber designed for optimal thermal conductance and for efficient optical viewing of reaction products in the chamber.

Abstract: The present invention provides methods, apparatuses and computer programs for verifying the integrity of a probe by comparing the fluorescence value of a probe to a threshold value. The invention also provides for methods, apparatuses and computer programs for normalizing the fluorescence value of a probe and detecting a target nucleic acid in a sample.

Abstract: A system for controlling the temperature of a reaction mixture comprises at least one heating device for heating the mixture and a power regulator for regulating the amount of power supplied to the heating device. A controller in communication with the power regulator includes program instructions for heating the reaction mixture by setting a variable target temperature that initially exceeds a desired setpoint temperature for the mixture. When the heating device reaches a threshold temperature, the variable target temperature is decreased to the desired setpoint temperature. In another embodiment, the controller includes an adaptive control program for dynamically adjusting the duration or intensity of power pulses provided to the heating device.

Abstract: The invention provides a device and method for the manipulation of materials (e.g., particles, cells, macromolecules, such as proteins, nucleic acids or other moieties) in a fluid sample. The device comprises a substrate having a plurality of microstructures and an insulator film on the structures. Application of a voltage to the structures induces separation of materials in the sample. The device and method are useful for a wide variety of applications such as dielectrophoresis (DEP) or the separation of a target material from other material in a fluid sample.

Abstract: The invention provides a multi-bead assay system for a protein based assay comprising at least two different beads. The first bead comprises protein and a protein stabilization matrix. The first bead forms a first solution when dissolved in liquid, and the first solution permits a first activity level for the assay. The second bead comprises a potentiation bead matrix that when dissolved in the first solution forms a second solution that potentiates the protein based assay to achieve a second activity level that is higher than the first activity level.

Abstract: Methods for balancing multiplexed PCR reactions are provided which exploit differences in primer and amplicon Tms. The methods may be controlled by a computer process. Also provided are articles of manufacture useful in such methods and compositions containing primers and probes useful in such methods.

Abstract: The present invention relates to methods and compositions providing optimized reaction conditions for nucleic acid amplification reactions and for methods and compositions that reduce contamination of amplification reactions during set up.

Abstract: An apparatus for disrupting cells or viruses comprises a container having a chamber for holding the cells or viruses. The container includes at least one flexible wall defining the chamber. The apparatus also includes a transducer for impacting an external surface of the flexible wall to generate pressure waves in the chamber. The apparatus also includes a pressure source for increasing the pressure in the chamber. The pressurization of the chamber ensures effective coupling between the transducer and the flexible wall. The apparatus may also include beads in the chamber for rupturing the cells or viruses.

Abstract: An apparatus for disrupting cells or viruses comprises a container having a chamber for holding the cells or viruses. The container includes at least one flexible wall defining the chamber. The apparatus also includes a transducer for impacting an external surface of the flexible wall to generate pressure waves in the chamber. The apparatus also includes a pressure source for increasing the pressure in the chamber. The pressurization of the chamber ensures effective coupling between the transducer and the flexible wall. The apparatus may also include beads in the chamber for rupturing the cells or viruses.

Abstract: An apparatus for disrupting cells or viruses comprises a container having a chamber for holding the cells or viruses. The container includes at least one flexible wall defining the chamber. The apparatus also includes a transducer for impacting an external surface of the flexible wall to generate pressure waves in the chamber. The apparatus also includes a pressure source for increasing the pressure in the chamber. The pressurization of the chamber ensures effective coupling between the transducer and the flexible wall. The apparatus may also include beads in the chamber for rupturing the cells or viruses.

Abstract: An apparatus for determining a threshold value (e.g., a threshold cycle number or a time value) in a nucleic acid amplification reaction comprises a detection mechanism for measuring, at a plurality of different times during the amplification reaction, at least one signal whose intensity is related to the quantity of a nucleic acid sequence being amplified in the reaction. A controller in communication with the detection mechanism is programmed to store signal values defining a growth curve for the nucleic acid sequence, determine a derivative of the growth curve, and calculate a cycle number or time value associated with a characteristic of the derivative.

Abstract: A container for holding cells or viruses for disruption comprises a chamber defined by two spaced apart, opposing major walls and side walls connecting the major walls to each other. At least one of the major walls has an external surface to which the transducer may be coupled and is sufficiently flexible to flex in response to vibratory motion of the transducer. The container also has at least one port for introducing the cells or viruses into the chamber. In some embodiments, the chamber contains beads for aiding the disruption of the cells or viruses.

Abstract: An analysis device comprises a body having a reaction chamber for chemically reacting a sample, a separation region for separating components of the sample, and a transition region connecting the reaction chamber to the separation region. The transition region includes at least one valve for controlling the flow of fluid between the reaction chamber and the separation region. Further, the transition region thermally isolates the reaction chamber from the separation region. In a preferred embodiment, the reaction chamber is an amplification chamber for amplifying nucleic acid in the sample, and the separation region comprises an electrophoresis channel containing a suitable matrix material, such as electrophoresis gel or buffer, for separating nucleic acid fragments. Electrodes are embedded in the body for separation of sample components. The body may also be surrounded by external, functional components such as an optical detector for detecting separated components of the sample.

Abstract: A method for determining an unknown starting quantity of a target nucleic acid sequence in a test sample comprises the steps of amplifying the unknown starting quantity of the target nucleic acid sequence in the test sample and known starting quantities of a calibration nucleic acid sequence in respective calibration samples; and determining a respective threshold value for each of the nucleic acid sequences using a derivative of a growth curve derived for the sequence. The starting quantity of the target nucleic acid sequence in the test sample is determined using the threshold value determined for the target sequence and a calibration curve derived from the threshold values determined for the known starting quantities of the calibration nucleic acid sequences. The invention also provides methods for determining a starting quantity of a nucleic acid sequence in a sample using quantitative internal controls or using internal standards.

Abstract: Described are methods, kits and systems for multiplexed detection of biological agents in a sample, e.g., multiplexed detection of e.g., bacteria, viruses, and biological toxins. The method utilizes two markers for each agent; the presence or absence in a sample of each of the two markers per agent is determined in separate reactions; however each reaction is used to detect a single marker for multiple agents. Also disclosed is the multiplexed detection method using real time PCR. The invention provides an efficient, cost-effective, and specific method for multiplexed detection of biological agents.

Abstract: A method for preparing a sample suspected to contain a target nucleic acid sequence for a nucleic acid amplification reaction and for verifying the effectiveness of the sample preparation comprises the step of mixing the sample with sample preparation controls. The sample preparation controls are cells, spores, microorganisms, or viruses that contain a marker nucleic acid sequence. The sample mixed with the sample preparation controls is subjected to a lysis treatment, and nucleic acid released by the lysis treatment is subjected to nucleic acid amplification conditions. The presence or absence of the target nucleic acid sequence and of the marker nucleic acid sequence is then determined. Positive detection of the marker nucleic acid sequence indicates that the sample preparation process was satisfactory, while the inability to detect the marker nucleic acid sequence indicates inadequate sample preparation.

Abstract: An apparatus for determining a threshold value (e.g., a threshold cycle number or a time value) in a nucleic acid amplification reaction comprises a detection mechanism for measuring, at a plurality of different times during the amplification reaction, at least one signal whose intensity is related to the quantity of a nucleic acid sequence being amplified in the reaction. A controller in communication with the detection mechanism is programmed to store signal values defining a growth curve for the nucleic acid sequence, determine a derivative of the growth curve, and calculate a cycle number or time value associated with a characteristic of the derivative.

Abstract: An analyte is separated from a fluid sample by introducing the sample into a cartridge having a sample port and a first flow path extending from the sample port. The first flow path includes an extraction chamber containing a solid support for capturing the analyte from the sample. The cartridge has a second flow path for eluting the captured analyte from the extraction chamber, the second flow diverging from the first flow path after passing through the extraction chamber. The sample is forced to flow through the extraction chamber and into a waste chamber, thereby capturing the analyte with the solid support as the sample flows through the extraction chamber. The captured analyte is then eluted from the extraction chamber by forcing an elution fluid to flow through the extraction chamber and along the second flow path.