Provided include methods, compositions, kits, and systems for replenishing a rolling circle amplification (RCA) reaction in a vessel. The RCA reaction can be initiated by contacting a nucleic acid template and a primer with a loading buffer comprising a DNA polymerase and polymerase extension agents including a divalent metal cation and a polyelectrolyte, followed by replenishing with an amplification buffer to continue the nucleic acid amplification through primer extension. The amplification buffer is different in composition from the loading buffer and does not comprise any DNA polymerase.

Provided include methods, compositions, kits, and systems for replenishing a rolling circle amplification (RCA) reaction in a vessel. The RCA reaction can be initiated by contacting a nucleic acid template and a primer with a loading buffer comprising a DNA polymerase and polymerase extension agents including a divalent metal cation and a polyelectrolyte, followed by replenishing with an amplification buffer to continue the nucleic acid amplification through primer extension. The amplification buffer is different in composition from the loading buffer and does not comprise any DNA polymerase.

The present disclosure is directed to aptamers comprising a detectable marker situated at an internal location within the aptamer, use of the aptamers to, e.g., detect target analytes, and methods of making the aptamers. In exemplary embodiments, methods of the disclosure comprise contacting the target analyte with an aptamer comprising a detectable marker situated at an internal location within the aptamer, wherein the contacting results in binding of the target analyte to the aptamer, wherein target analyte binding to the aptamer results in restriction of internal rotation of the marker, resulting in a detectable change in the marker.

The present disclosure is directed to aptamers comprising a detectable marker situated at an internal location within the aptamer, use of the aptamers to, e.g., detect target analytes, and methods of making the aptamers. In exemplary embodiments, methods of the disclosure comprise contacting the target analyte with an aptamer comprising a detectable marker situated at an internal location within the aptamer, wherein the contacting results in binding of the target analyte to the aptamer, wherein target analyte binding to the aptamer results in restriction of internal rotation of the marker, resulting in a detectable change in the marker.

Provided herein are methods of amplifying and detecting pathogens and drug resistance markers (e.g., antibiotic resistance genes) in complex samples (e.g., blood), as well as related panels and compositions (e.g., primers, probes, magnetic particles, systems, cartridges, and kits). The methods, panels, and compositions can be used for comprehensive detection of pathogens and drug resistance markers for patient identification, patient selection, optimization of therapies, and antimicrobial stewardship.

Provided herein are methods of amplifying and detecting pathogens and drug resistance markers (e.g., antibiotic resistance genes) in complex samples (e.g., blood), as well as related panels and compositions (e.g., primers, probes, magnetic particles, systems, cartridges, and kits). The methods, panels, and compositions can be used for comprehensive detection of pathogens and drug resistance markers for patient identification, patient selection, optimization of therapies, and antimicrobial stewardship.

The present disclosure describes a method for detecting the presence of Mycobacterium tuberculosis in a bodily fluid sample. The method utilizes CRISPR effector proteins along with a guide RNA and a reporter molecule, such that when the guide RNA hybridizes with a target nucleotide fragment, the CRISPR effector protein cleaves the reporter molecule, resulting in a detectable signal.

The present disclosure describes a method for detecting the presence of Mycobacterium tuberculosis in a bodily fluid sample. The method utilizes CRISPR effector proteins along with a guide RNA and a reporter molecule, such that when the guide RNA hybridizes with a target nucleotide fragment, the CRISPR effector protein cleaves the reporter molecule, resulting in a detectable signal.

The present disclosure discloses a real-time fluorescence quantitative polymerase chain reaction (PCR) detection method and kit based on a metal ruthenium complex. The present disclosure is capable of establishing the detection method for performing a real-time fluorescence quantitative PCR by using the metal ruthenium complex as fluorescence dye.

The present disclosure discloses a real-time fluorescence quantitative polymerase chain reaction (PCR) detection method and kit based on a metal ruthenium complex. The present disclosure is capable of establishing the detection method for performing a real-time fluorescence quantitative PCR by using the metal ruthenium complex as fluorescence dye.