Molecular Diagnostics Technologies

LLNL scientists have designed a rapid PCR technology that incorporates the use of microfluidic thermal heat exchanger systems and is comprised of a porous internal medium, with two outlet channels, two tanks, and one or more exchanger wells for sample receiving. The wells and their corresponding inlet channels are coupled to two tanks that contain fluid with cold and hot temperatures. A controller is used to dictate the position of the fluid pump’s valves, which directs fluid flow between tanks. The fluid passes though the system’s porous medium, heating or cooling the samples being housed in the wells. When the fluid passes through the matrix, it provides extremely fast heat conduction that enables rapid thermal transfer between the fluid, matrix, and sampler holder.

LLNL has developed a new technology that provides a method for near-instantaneous heating of aqueous samples in microfluidic devices. The technology relates to a heating method that employs microwave energy absorption from a coincident low power Co-planar waveguide or microwave microstrip transmission line embedded in a microfluidic channel to instantaneously heat samples. The method heat samples in a focused area within a microfluidic channel on miniaturized chips. Aqueous solution microwave heating allows extremely fast heat transfer for both heating and cooling. This method/device provides a major advantage over current heating methods such as joule-heating from trace resistors which are time-consuming and provide an associated whole device heat build-up.

This LLNL-developed invention is multiplexed and utilizes the Luminex bead-based liquid array, which contains 100 different unique beads. Oligonucleotide probes with sequences complementary to the target sequences are covalently coupled to these unique beads. These capture beads are mixed with viral samples obtained from the patient via cheek swabbing or a throat wash and subjected to PCR in a conventional thermocycler. The amplified target sequence is then hybridized to complementary capture oligonucleotide probes via forward biotinylated primers. If this bead-probe-amplicon unit contains the target nucleic acid, it will be bound by the reporter molecule and fluorescence will be detected by a flow cytometer. This multiplexed assay would thus be able to detect and identify respiratory…

Researchers at LLNL have developed an instantaneous sample heating method to efficiently deposit thermal energy into a continuous stream or segmented microdroplets on a MOEMS device using an optimally low energy, commercially available CO2 laser. The device uses an ideal wavelength (absorption in the far infra-red (FIR) region (λ=10.6 μm)) to instantaneously heat fluidic partitions. The wavelength is absorbed by water molecules and waste little energy because, unlike typical PCR heating elements, the device itself is not heated by the laser. Instead the aqueous solution directly absorbs the heat. This technology is a major improvement over current microfluidic channel heating methods.

Researchers at LLNL have designed a new technology that allows the integration of a large bench-top thermal cycling instrument onto a miniaturized instrument. This instrument is powered and controlled by portable thumb-drive systems such as an USB. USB thumb-drives are commonly used to transfer data from the instrument onto a PC, however, in this new technology the thumb drive becomes the instrument itself! LLNL researcher’s technology includes thermocycling configured for low power and efficiency, miniaturization of components and controllers, fabrication on a solid-state thumb drive, and integration with USB data and supplied power. This system uses bus power for thermal cycling and bus data lines for data transmission and programming, which allows for portable power.

LLNL researchers have developed a high-volume, low-cost diagnostic test that is easy to use and provides results in under an hour. The testing platform will provide emergency responders and other medical professionals with the ability to screen individuals using oral and nasal samples, and obtain results in approximately 30 minutes. This point-of-care testing approach will enable rapid triage of a high volume of patients, without needing to send a sample to a laboratory for testing and then waiting for results. The easy-to-use, compact testing kit consists of a single, disposable tube, which is used throughout the process to collect samples from patients and obtain a positive or negative test result.

This technology describes a method for performing immediate in-line sample heating to promote the required chemical reactions for amplification, activation, or detection, depending on the thermodynamics of the particular assay involved. The basis of this technology is a method that employ microwave energy absorption to instantaneously heat fluidic partitions without heating the device itself or any oil, or entrapped air. With this invention little energy is wasted heating the device and instead is absorbed heating the aqueous solution within the microfluidic device’s chambers, channels, or reservoirs. This will allow the most efficient, fastest, and best method for energizing chemical reactions in microfluidics devices.

LLNL researchers have developed a new method for faster, more accurate, and precise thermal control for DNA amplification. This technology uses sensor-controlled nodes to monitor and cycle materials through a microfluidic heat exchanging system. Thermal energy travels from a power module through thermal electric elements to sample wells. Sensors coupled to each sample well monitor and respond to predetermined temperature thresholds allowing for the simultaneous directional transfer of thermal energy and therefore better thermal cycling controls. When using LLNL’s solid-state distributed node-based rapid thermal cycler, researchers can be assured that sample DNA is being amplified under optimal conditions.

LLNL scientists have developed a technology which fulfills this need. The LLNL technology itself is comprised of two elements which are to be embedded in a user's personal electronic device (e.g. cell phone, tablet device, pager, etc.). The first is a proximity monitor which transmits location and temporal data such as the distance between the user and a contagious individual and the duration of proximity. The second is a personal exposure notification which comes after the user has been positively diagnosed with a contagious disease by a healthcare provider. Information of their contagiousness is downloaded to a server and the user's device would then transmit exposure warnings to other individuals who have encountered the user and are deemed by the proximity monitor to be at high…

LNLL scientists have invented a method for multiplexed detection of PCR amplified products which can be completed in a single step. Highly validated species-specific primer sets are used to simultaneously amplify multiple diagnostic regions unique to each individual pathogen. Resolution of the mix of amplified products is achieved by PCR product hybridization to corresponding probe sequences, attached to unique sets of fluorescent beads in liquid. The hybridized beads are processed through a flow cytometer, which detects presence and quantity of each PCR product. The assay is optimized to allow for maximum sensitivity in a multiplexed format. A background PCR product is formed via background multiplex PCR amplification reaction using a control DNA sequence. Comparing the fluorescence…


LLNL scientists have developed a rapid parallel genetic profiling technology that can be used to detect an array of pathogens from a small, complex sample. Detectable pathogens by the LLNL technology include viruses, bacteria, protozoa, and other microbes. The device works by first splitting a given sample into millions of emulsified, encapsulated microdroplets each of which are then split once more and run through a parallel analysis consisting of both a genomic and a proteomic assay. The droplets within the assays are first run through a PCR process which amplifies even the smallest quantities of DNA or RNA for analysis. Finally, the droplets, now with sufficient quantities of DNA or RNA for analysis after PCR, are dissolved and run on an agarose gel via electrophoresis. Analysis of…

LLNL researchers have developed a method to quickly and accurately identify the family of a virus infecting a vertebrate via PCR. Universal primer sets consisting of short nucleic acid strands of 7 to 30 base pairs in length were created to amplify target sequences of viral DNA or RNA. These primers can amplify certain identifying sequences of all viral genomes sequenced to date as well as numerous virus subgroupings. The PCR products are separated on a gel by gradient electrophoresis to identify the virus. Altogether, these primers can identify all 28 known virus families that affect vertebrates. Each strain or species of virus produces its own unique electrophoretic banding signature, allowing for easy and quick virus identification. Primer libraries will be updated as new virus…


LLNL researchers have invented a system for identifying all known and unknown pathogenic or non-pathogenic organisms in a sample. This invention takes a complex sample and generates droplets from it. The droplets consist of sub-nanoliter volume reactors which contain the organism sized particles. A lysis device lyses the organisms and releases the nucleic acids. An amplifier then magnifies the quantity of available nucleic acids. Then, a fractionator liberates the nucleic acids from the droplets. Finally, a parallel analyzer identifies all the known and unknown pathogen or non-pathogenic organisms in the complex sample. This device functions with DNA or RNA samples.


LLNL scientists have created a standalone pathogen identifier that can be placed in public settings, such as in stores or on street corners. Not unlike an ATM in physical size, this kiosk will accept biological samples from an individual for multiplexed analysis. The sample collection process will be sufficiently simple such that anyone could begin the diagnostic process after making the appropriate payment via cash, credit, or debit cards. After the customer signs the appropriate disclaimers concerning diagnosis and liability, a sterile swab or collection tool or vial, viral transport media, instructions on collecting a sample, gloves, and antiseptic wipes would be dispensed to them. The customer then would select what pathogens they want to be screened for before the assay begins.…

The LLNL invention has two assay chambers wherein each chamber is comprised of another two chamber modules. This allows the device to process up to two assays per chamber module, or four total assays per biological sample. These two duplex assays are each fed by parallel interrogation ports while the device still maintains a small physical profile. Each port has its own LED for excitation, allowing the second assay to have particularly improved excitation. The excitation of both assay chambers is achieved by a much simplified and straightforward optical system, which lowers the size, complexity, and cost of the device while increasing reliability and performance.


LLNL researchers have developed a portable device which analyzes one or multiple types of body fluids or gases to test for one or more medical conditions. A bodily fluid (such as blood, perspiration, saliva, breath, or urine) is put into a condenser surface and is then separated into both a primarily gas fluid component and a second one that is primarily liquid. These two samples from the same fluid or gas source are subjected to analysis by, in various combinations, five different instruments: a condenser, functionalized nanostructures, an optional volatilizer, a differential mobility spectrometer, and an optional biomarker analyzer. Each instrument provides a unique analysis of a physical or chemical element of the tested bodily fluid.