Brandeis University

Other

Kenneth E Pierce

Lawrence J Wangh

John E Rice

Cristina Hartshorn

J Aquiles Sanchez

Arthur Henry Reis, Jr

Jesse J Salk

Arthur Reis

Skye Fishbein

Stephen Van Hooser

Adam E Osborne

Alexandra Isabel King Over

Mark Levey

J Acquiles Sanchez

Jesus A Sanchez

Provided herein are reagents and kits for detection of multiple target sequences in a single-tube, single-color assay, and methods of use thereof. In particular, multiplex assays are provided for the detection ofcomplex target sequences (e.g., katG, rpoB, inhA promotor, pncA, etc.).

Multiplex target detection assay

Provided herein are reagents for improving PCR accuracy.

Reagents for improving PCR accuracy

Provided herein are mispriming prevention reagents, compositions and kits comprising such reagents and methods of use thereof.

Mispriming prevention reagents

The present invention relates to compositions and methods for nucleic acid based diagnostic assays. In particular, the present invention provides primers for asymmetric PCR and other amplification modalities. In some embodiments, the present invention provides multiple primers for amplification of related nucleic acid targets in a single reaction.

Compositions and methods for nucleic acid based diagnostic assays for variable sequence targets

Homogenous detection during or following PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled primers and probes, improves reproducibility and quantification. Low-temperature homogeneous detection during or following non-symmetric PCR amplification, preferably LATE-PCR, utilizing fluorescent DNA dye and indirectly excitable labeled mismatch-tolerant probes permits analysis of complex targets. Sequencing sample preparation methods following LATE-PCR amplifications reduce complexity and permit 'single-tube' processing.

Methods for sequential DNA amplification and sequencing

Provided herein are methods, kits, and compositions related to nucleic acid detection assays that allow discrimination of multiple target sequences with a single probe. In particular, provided herein are methods kits, and compositions that include single-probe target sequence discrimination where different target amplicons may have identical probe hybridization sequences by employing multiple temperature end-point signal probe detection. Also provided herein are methods, kits, and compositions for distinguishing between two or more target amplicons using multiple-temperature end-point probe detection. In certain embodiments, asymmetric PCR amplification methods are employed (e.g., LATE-PCR amplification).

Single probe, multiple temperature, nucleic acid detection methods, kits, and compositions

A non-symmetric polymerize chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25&#xb0; C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

Late-PCR

Provided herein are methods, kits, and compositions related to nucleic acid detection that allow the detection and discrimination of variousspecies and types. In particular, provided herein are assays that allow the detection of MSSA, MSSE, MRSA, MRSE, VRSA, VRSE, and MRSA community strain by detecting the presence of absence of one or more target sequences selected from a list of target genes and sequences that include, but are not limited to: mecA, femA-SA, femA-SE, lukF-PV, lukS-PV, VanA, SCCmec type I, type II, type III, type IV, type V, and type VII. In certain embodiments, asymmetric PCR amplification methods are employed. In other embodiments, probe-target generated signals are detected at least two temperatures to generate a temperature/temperature signal ratio.

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A lightweight tender comprising a hull constructed from substantially a single piece of composite material and a retractable pillar attached to the hull, wherein the retractable pillar defines a longitudinally extending channel. A retractable carbon fiber bimini cover is pivotingly attached to the retractable pillar, and defines a top deck when the pillar is retracted, and defines a bimini top extended. A first end of a brace is hingedly attached to the bimini cover and a second end of the brace travels within the longitudinally extending channel of the retractable pillar such that the angle between the bimini cover and the retractable pillar increases as the brace travels distally within the longitudinally extending channel.

Composite tender with retractable bimini hard-top

A non-symmetric polymerise chain reaction (PCR) amplification method employing a limiting primer in low concentration whose concentration-adjusted melting point at least equals, and preferably exceeds, that of the excess primer, the latter in turn not being more than 25&#xb0; C. below the melting temperature of the amplicon. Assays employing such amplification and labeled hybridization probes, including assays that include a detection step following primer extension or a low-temperature probe, or both. Kits for performing such assays and primer or primer-and-probe sets for performing the foregoing amplifications and assays.

An assay comprising more than one primer pair and more than one detection probe, a low copy number synthetic amplicon corresponding to each of the primer pairs. The assay can detect and distinguish between various sub-types and strains of an influenza virus using any suitable nucleic acid amplification technique. Related kits and methods are also described.

Detection and analysis of influenza virus

Primers, probes and methods for nucleic acid amplification

An additive for preventing mispriming in polymerase chain reaction (PCR) amplifications and assays comprising a hairpin oligonucleotide having a stem duplex greater than six nucleotides in length and a stabilized stem terminus. The additive improves PCR amplifications, including LATE-PCR amplifications when added to initial amplification reaction mixtures. It can be included in oligonucleotide sets and in kits for PCR amplification and assays.

Growing community of inventors

Natick, MA, United States of America

Kenneth E Pierce