Embodiments of the disclosure encompass methods of amplifying nucleic acid from one or more cells using MALBAC (multiple annealing and looping-based amplification cycles) primers. In particular embodiments, the nucleic acid is amplified as amplicons in a linear manner. Specific embodiments include the removal or effective destruction of nonlinearly produced amplicons.

The invention generally relates to assemblies for displacing droplets from a vessel that facilitate the collection and transfer of the droplets while minimizing sample loss. In certain aspects, the assembly includes at least one droplet formation module, in which the module is configured to form droplets surrounded by an immiscible fluid. The assembly also includes at least one chamber including an outlet, in which the chamber is configured to receive droplets and an immiscible fluid, and in which the outlet is configured to receive substantially only droplets. The assembly further includes a channel, configured such that the droplet formation module and the chamber are in fluid communication with each other via the channel. In other aspects, the assembly includes a plurality of hollow members, in which the hollow members are channels and in which the members are configured to interact with a vessel. The plurality of hollow members includes a first member configured to expel a fluid immiscible with droplets in the vessel and a second member configured to substantially only droplets from the vessel. The assembly also includes a main channel, in which the second member is in fluid communication with the main channel. The assembly also includes at least one analysis module connected to the main channel.

Provided herein are methods of reliably determining whether or not an individual is a user of tobacco.

The invention concerns nucleic acids coding for mutated or truncated forms of the human parkin gene, or forms comprising multiplication of exons, and the corresponding proteins and antibodies. The invention also concerns methods and kits for identifying mutations of the parkin gene, and for studying compounds for therapeutic purposes.

The present invention refers to a process for classifying tumor samples of unknown and/or uncertain primary origin, specifically including the steps of obtaining patterns of biological activity modulation of tumor of unknown and/or uncertain primary origin and comparing them to an specific and unique group of biomarkers which determine the profiles of biological activity modulation of known origin tumors. The present invention belongs to the molecular biology and genetics field.

The present invention relates to methods for predicting the survival time of patients suffering from cancer. Said methods are based on the quantification and analysis of the cell free nucleic acids that are present in a sample from the patient and typically include the determination of the level of the mutant nucleic acid which contains a mutation of interest, the calculation of the mutation load for said mutation of interest, the calculation of the DNA integrity index or a combination thereof.

Disclosed are methods and compositions for detecting the presence of a cancer in a subject and assessing the efficacy of treatments for the same. The disclosed method use reverse transcription polymerase chain reaction (RT-PCR) and multiplex polymerase chain reaction techniques as well as Template Exchange Extension Reaction (TEER) to detect the presence of point mutations, truncations, or fusions of anaplastic lymphoma kinase.

A method of detecting a cancer progression observation index gene group, comprising: a) preparing a plurality of AKR/J mice in which an oncogene is inserted into a thymocyte; b) dividing the AKR/J mice into three groups, raising the first group of the AKR/J mice after high-dose ionizing radiation of 0.8 Gy/min, raising the second group of the AKR/J mice after low-dose ionizing radiation of 0.7 mGy/hr, and raising the third group of the AKR/J mice in a general environment; c) obtaining a thymus of a dead mouse of the first or second group of the AKR/J mice throughout the b) operation and diagnosing cancer when a weight of the thymus is increased to twice or more than before radiation; d) extracting thymuses by sacrificing the first to third groups of the AKR/J mice at the time at which the third group of the AKR/J mice initially die; and e) selecting only a thymus having no change in weight compared to the organs of the same-aged non-irradiated AKR/J mice from the organs extracted in d) and detecting a gene of the organ whose weight is increased or decreased by a factor of two or more through gene analysis.

Compositions and methods for detecting Trichomonas vaginalis are provided.

The invention relates to the field of virology. The invention provides an isolated essentially mammalian negative-sense single-stranded RNA virus (MPV) within the subfamily Pneumovirinae of the family Paramyxoviridae and identifiable as phylogenetically corresponding to the genus Metapneumovirus and components thereof.

A microbial gaseous-fluid sampler for collecting microbial particles from gaseous fluid includes a gaseous-fluid intake portion having a sample head with a plurality of holes. The gaseous-fluid intake portion further includes a collar configured to receive a Petri dish including agar. The plurality of holes define an exit plane that is positioned a distance from the agar within a range of 5.5 millimeters to 7.5 millimeters. The velocity of the air exiting the plurality to holes is within a range of 18.5 meters per second to 20.5 meters per second.

In some embodiments, the present disclosure pertains to a method for retrieving at least one molecular recognition element in a fixed tissue. In some embodiments the method comprises preparing a solution comprising at least one aldehyde-scavenging agent. In some embodiments, the method comprises contacting the fixed tissue with the solution. In some embodiments, the tissue is fixed with an aldehyde-based cross-linking agent. In some embodiments, a reaction of the aldehyde-scavenging agent with the aldehydes comprising the cross-linking agent retrieves the at least one molecular recognition t. In some embodiments, the at least one rolecular recognition element comprises of amino acids, peptides, proteins, nucleic acids, carbohydrates, lipids, or a combination thereof. In some embodiments, the at least one aldehyde-scavenging agent comprises of beta-dicarbonyl compounds, mono or di-amide scavengers, ethyl alcohols, sulfur containing compounds, mercaptoethylamines, mercaptoethanols, hydrazines, ethanolamines, hydroxylamines, anilines, variation of amines, activated charcoal, phenols, or mixtures and combinations thereof.

The present disclosure relates to use of high-concentration aldehyde-based fixatives to fix tissue samples. Aldehyde concentrations are selected that significantly increase rate of diffusion of the fixative solution into the tissue. When combined in the cold-temperature step of a two-temperature fixation, a substantial improvement in the preservation of post-translationally modified proteins is achieved.

Highly efficient and rapid filtration-based concentration devices, systems and methods are disclosed with sample fluidic lines and a filter packaged in a disposable tip which concentrate biological particles that are suspended in liquid from a dilute feed suspension. A sample concentrate or retentate suspension is retained while eliminating the separated fluid in a separate flow stream. The concentrate is then dispensed from the disposable tip in a set volume of elution fluid. Suspended biological particles include such materials as proteins/toxins, viruses, DNA, and/or bacteria in the size range of approximately 0.001 micron to 20 microns diameter. Concentration of these particles is advantageous for detection of target particles in a dilute suspension, because concentrating them into a small volume makes them easier to detect. All conduits by which the disposable tip attaches to the instrument are combined into a single connection point on the upper end of the tip.

A system and method for concurrently and uniformly removing thermal energy from clustered specimen samples.

In flow cytometry, particles (2) can be distinguished between populations (8) by combining n-dimensional parameter data, which may be derived from signal data from a particle, to mathematically achieve numerical results representative of an alteration (48). An alteration may include a rotational alteration, a scaled alteration, or perhaps even a translational alteration. Alterations may enhance separation of data points which may provide real-time classification (49) of signal data corresponding to individual particles into one of at least two populations.

Aspects of the present disclosure include methods for processing a biological sample. Methods according to certain embodiments include contacting a biological sample having cells with an assay reagent that includes one or more analyte-specific binding members to produce a biological sample assay composition and introducing the biological assay composition into an inlet of a flow cytometer having an integrated acoustic separator. Systems, including a flow cytometer with integrated acoustic separator having one or more acoustic concentrator devices suitable for practicing the subject methods are also described.

The invention is drawn to novel nanostructures comprising hollow nanospheres and nanotubes for use as chemical sensors, conduits for fluids, and electronic conductors. The nanostructures can be used in microfluidic devices, for transporting fluids between devices and structures in analytical devices, for conducting electrical currents between devices and structure in analytical devices, and for conducting electrical currents between biological molecules and electronic devices, such as bio-microchips.

The present invention provides methods of determining cell sensitivity to a therapeutic agent.

The present invention provides a method for imaging microtentacles on isolated, living, non-adherent primary tumor cells from a cancer subject comprising: i) obtaining one or more living, non-adherent primary tumor cells that has been isolated from a solid tumor from the subject; and ii) imaging the one or more living, non-adherent primary tumor cells and detecting the microtentacles.

An assay and a method for identifying a prebiotic to treat a subject in need thereof to promote intestinal barrier integrity or to blunt an inflammatory response are provided.

Light harvesting luminescent multichromophores that are configured upon excitation to transfer energy to, and amplify the emission from, an acceptor signaling chromophore in energy-receiving proximity therewith are provided. Also provided are compositions for labelling a target. The labelling composition may include a donor light harvesting multichromophore and an acceptor signaling chromophore in energy-receiving proximity to the donor light harvesting multichromophore. Also provided is an aqueous composition for labelling a target, including: a donor light harvesting multichromophore; an acceptor signaling chromophore in energy-receiving proximity therewith; and a sensor biomolecule. Methods for using the subject compositions are also provided.

The present invention relates to a novel method for detecting a detection object in a sample, and a detection device using the same. The detecting method of the present invention uses a “bridge composite” in which gold nanoparticles and an antibody specific to a detection object are coupled in order to induce a sufficient coupling reaction between the antibody and the detection object, thereby improving reactivity. Accordingly, since excellent resolution is provided, the method of the present invention has advantages of enabling accurate concentration measurement of a detection object in a sample, and amplifying a measurement signal. In addition, the method of the present invention can effectively detect small molecules such as hormones, vitamins, etc. having a small molecular weight.

Methods of screening for expression of an RNA associated with a post-kidney transplant complication include collecting vesicles from urine, isolating vesicle-associated RNA, and analyzing expression patterns. In particular, AIF1, BTN3A3, CCL5, CD48, HAVCR1, or SLC6A6 mRNA expression patterns are analyzed.

A sensing sensor includes a wiring board, a piezoelectric resonator, a channel forming member, a case body, and a regulating portion. The case body houses the wiring board, the piezoelectric resonator, and the channel forming member. The case body includes a window faced to a region including the output terminal on the another surface side of the wiring board and an injection port supplying a supply liquid to the one end side of the channel. The regulating portion is disposed in the case body. The regulating portion regulates the wiring board from the one surface side when the wiring board is pressed from the another surface side. The piezoelectric resonator is pressed against the surrounding portion by the output terminal being pressed from the another surface side.

An assay system and method for use in the field of chemical testing is disclosed. The assay system can be used for filtering whole blood for testing on an integrated circuit containing digital control functionality.

Multi-biotinylated reactants are provided which can be used in divalent complexes for various applications such as colocalization, labeling, immobilization, and purification. Methods for constructing, purifying, and using the bis-biotinylated reactants are also provided. In certain embodiments, two bis-biotinylated reactants are bound to a single streptavidin tetramer to provide a complex having a 1:1 stoichiometry with respect to the bis-biotinylated reactants.

Viruses and other bioagents are of high medical and biodefense concern and their detection at concentrations well below the threshold necessary to cause health hazards continues to be a challenge with respect to sensitivity, specificity, and selectivity. Ideally, assays for accurate and real time detection of viral agents and other bioagents would not necessitate any pre-processing of the analyte, which would make them applicable for example to bodily fluids (blood, sputum) and man-made as well as naturally occurring bodies of water (pools, rivers). We describe herein a robust biosensor that combines the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In preferred embodiments, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV), a member of the genus Hantavirus, family Bunyaviridae, negative-stranded RNA viruses. Rapid detection (within seconds) of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, although the sensor was approximately 50×104-fold more sensitive for the detection of SNV. For both pathogens, the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1. The biosensor was able to detect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS). Further, in a proof-of-principle real world application, the SAW biosensor was capable of selectively detecting SNV agents in complex solutions, such as naturally occurring bodies of water (river, sewage effluent) without analyte pre-processing.

Methods and compositions are described for categorizing and treating autoimmune disease, using single cell network profiling (SCNP), where activation levels of one or more activatable elements are determined in single cells, with or without modulation, to categorize or determine treatment for the autoimmune disease.

The present invention provides assays for detecting and measuring the presence or level of anti-TNFα drug therapeutics and autoantibodies in a sample. The present invention is useful for optimizing therapy and monitoring patients receiving anti-TNFα drug therapeutics to detect the presence or level of autoantibodies (e.g., HACA and/or HAHA) against the drug.

An antigenic characterization method using polyclonal antibody-based proximity ligation assays (polyPLA). Methods, kits, and other tools disclosed herein are useful in detecting microbial antigenic variants in samples, including clinical samples. The methods and kits have great utility in detecting antigenic variants for pathogenic microbes, including viruses, bacteria, and parasites.

The invention concerns soluble and antigenic HTLV p24 variants that can be fused to chaperones and their use in diagnostic applications such as immunoassays for detecting antibodies against HTLV-I or HTLV-II in an isolated biological sample. In particular, the invention relates to a soluble HTLV-I or HTLV-II p24 antigen comprising either the N- or the C-terminal domain of p24 and lacking the other domain. Moreover, the invention covers recombinant DNA molecules encoding these HTLV-I and -II fusion antigens as well as their recombinant production using expression vectors and host cells transformed with such expression vectors. In addition, the invention focuses on compositions of these HTLV p24 antigens with HTLV gp21 antigen and on an immunoassay method for detection of HTLV antibodies using the antigens of the invention. Also the use of HTLV p24 antigens in an in vitro diagnostic assay as well as a reagent kit for detection of anti-HTLV-antibodies comprising said HTLV antigens is encompassed.

This invention publicizes a chronic renal disease diagnostic kit, comprised of FGF23 monoclonal antibody, biotin labeling reagent Sulfo-NHS-LC-Biotin, PBS buffer solution (pH7.0, 0.1M), casein saline solution, streptomycin avidin and substrate TMB. The labeling condition is the weight ratio of FGF23 antibody and Sulfo-NHS-LC-Biotin, which is 1:5-1:12. The dilution ratio of streptomycin avidin is 1:3000. The lowest testing threshold limit value of FGF23 kit is 10 pg/ml, whose sensitivity is 10 times higher than that of common monoclonal antibody, which is very significant for early diagnose of chronic renal disease.

The present invention provides methods, compositions and kits for the characterization of cellular pathways in cells containing genetic alterations.

The present invention relates to in vitro methods of assessing the susceptibility or responsiveness of a subject to the treatment with an epidermal growth factor receptor (EGFR) inhibitor/antagonist, wherein the subject has been diagnosed or suspected of suffering from inflammation-associated. These methods comprise determining the level of expression of EGFR in myeloid cells in a sample from the subject, wherein an expression of EGFR in the myeloid cells is indicative of the subject being susceptible to the treatment with an EGFR inhibitor/antagonist. The invention also relates to EGFR inhibitors/antagonists for use in the treatment or amelioration of inflammation-associated cancer. The invention furthermore provides in vitro methods of prognosing the survival time, progression-free survival time or disease course of a subject that has been diagnosed or suspected of suffering from from inflammation-associated cancer. In addition thereto, the invention relates to in vitro diagnostic methods of assessing the proneness of a subject to develop inflammation-associated cancer in which the expression of EGFR is determined in myeloid cells from the subject.

The present invention provides an approach for the determination of the activation states of a plurality of proteins in single cells. This approach permits the rapid detection of heterogeneity in a complex cell population based on activation states, expression markers and other criteria, and the identification of cellular subsets that exhibit correlated changes in activation within the cell population. Moreover, this approach allows the correlation of cellular activities or properties. In addition, the use of modulators of cellular activation allows for characterization of pathways and cell populations. Several exemplary diseases that can be analyzed using the invention include AML, MDS, and MPN.

The present invention relates to an in vitro method of diagnosing chronic myelomonocytic leukemia (CMML) in a subject, said method comprising the steps of: a) Detecting a monocyte population in a biological sample from said subject; b) Quantifying the CD14+/CD16− monocytes in said biological sample; c) Comparing the value of step b) to a reference value; and d) Diagnosing CMML based on said comparison. Preferably, said detecting step a) is performed by an exclusion gating strategy by flow cytometry.

A method to detect prostate cancer comprising contacting a sample of prostate cells from the patient with a set of detectably labeled probes under hybridization conditions and determining the presence of chromosomal abnormalities in prostate tumor tissue, PIN (intra-epithelial neoplasia), histologically benign tissue and benign prostatic hyperplasia (BPH); a method to combine immunofluorescence and FISH (IF-FISH) to facilitate the assessment of chromosomal abnormalities; a set of probes; and a kit comprising the set of probes and instructions for diagnosing prostate cancer in a patient.

We have discovered a protein in humans, herein referred to as collagen like gene (CLG) product (SEQ ID NOS: 12 and 13), that is expressed in human prostate cancer and breast cancer cell lines but not in normal adult, placenta, lung, liver, skeletal muscle, kidney or pancreas tissues. We have also discovered that the level of CLG mRNA expression correlates positively with the metastatic potential of the cancer cell lines tested.

The present invention provides detection methods for detecting a pre-cancerous epithelial cell signature. The present invention further provides reagents for use in the detection methods. A subject detection method is useful in various imaging, diagnostic, prognostic, and patient monitoring methods, which are also provided.

The diagnosis of male infertility is based predominantly on the results of standard semen analysis for concentration, total motility, progressive motility, volume, pH, viscosity and/or morphology. When sperm enter the female reproductive tract, they must undergo a series of physiological changes, known as capacitation, in order to fertilize an egg. This process involves plasma membrane changes that occur in response to stimuli within the female tract. These changes include removal of sterols and redistribution of the ganglioside GM1. Semen analysis identifies only half the cases of male infertility due to standard semen analysis providing little information on sperm functional competence. Previous data demonstrated that localization of the ganglioside, GM1, identifies sub-populations of sperm capable of undergoing the functional maturation process known as capacitation and tracks strongly with fertility.

Methods for multiplexed delivery of agents to a solid tissue in vivo followed by assessment of efficacy with mass spectrometry are described.

A system and method for detecting a biomarker in exhaled breath condensate nanodroplets comprises noninvasively collecting exhaled breath condensate nanodroplets of a subject, and analyzing said nanodroplets utilizing immuno-quantitative polymerase chain reaction to detect one or more target biomarkers.

The present invention relates to methods of determining if a subject has an increased risk of suffering from memory impairment. The methods comprise analyzing at least one sample from the subject to determine a value of the subject's exosomal profile or combined biomarker profile (lipids plus exosomal cargo) and comparing the value of the subject's exosomal or combined biomarker profile with the value of a normal exosomal or biomarker profile, respectively. A change in the value of the subject's exosomal or combined biomarker profile, including a change in the subject's exosomal or combined biomarker profile, over normal values is indicative that the subject has an increased risk of suffering from memory impairment compared to a normal individual.

In sporadic Alzheimer's disease, neurofibrillary lesion formation is preceded by extensive post-translational modification of the microtubule associated protein tau. Immunoassays have been developed recently that detect tau in biological specimens, thus providing a means for pre-mortem diagnosis of Alzheimer's disease, which has remained elusive. These assays have been improved by the analysis of relevant post-translational modifications, such as phosphorylation, however opportunity for improvement remains. The present invention addresses this issue by disclosing synthetic methylated peptides derived from the tau protein of paired helical filaments and non-diseased control brain. Alzheimer's disease specificity is provided by the presence or absence of methyl moieties on lysine residues and differences between mono-, di-, and tri-methylation. The methylated peptide is useful as an antigen and a binding partner for identifying compounds that interact with the peptide and the methylated tau protein, including antibodies that can distinguish non-diseased brain from that affected by Alzheimer's disease. The resulting antibodies are useful diagnostically and therapeutically. The compounds that specifically bind to methylated tau proteins are useful for eliminating abnormally methylated tau.

Provided are a detection agent for detecting hydroxy vitamin D, preparation method thereof, and use thereof in 25-hydroxy vitamin D immunological detection. The detection agent comprises a conjugate formed by a 25-hydroxy vitamin D antigen derivative and protein carrier, and magnetic spheres coated by the conjugate. Also provided is a 25-hydroxy vitamin D detection kit comprising the detection agent.