Direct amplification and detection of viral and bacterial pathogens
AssigneeQuest Diagnostics Investments Incorporated
InventorsMaurice Exner, Luca Jacky, Yin-Peng Chen, Huong Mai, Michelle M. Tabb, Michael Aye
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Abstract
Provided are methods for identifying the presence or absence of a target nucleic acid from a microorganism using direct amplification without a step of extraction of the nucleic acids, but retaining substantially the same specificity and sensitivity of methods assaying extracted nucleic acids. Further provided are reagent mixtures that allow for direct amplification of a sample, without the step of nucleic acid extraction.
Claims28
A method for identifying the presence or absence of a target nucleic acid from a microorganism in a biological sample obtained from a human, said method comprising:
- (a) contacting the sample with a DNA polymerase and a buffer under conditions suitable for amplification of the target nucleic acid from the sample without extracting the target nucleic acid from the sample;
- (b) thermocycling the sample from step (a) such that the target nucleic acid, if present, is amplified;
- and (c) detecting the amplified target nucleic acid, if present, produced from step (b), wherein nucleic acid in the sample is not extracted from the sample prior to amplification, and wherein said buffer comprises a cationic surfactant.
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A method for identifying the presence or absence of a target nucleic acid from a microorganism in a biological sample obtained from a human, said method comprising:
- (a) contacting the sample with a DNA polymerase and a buffer under conditions suitable for amplification of the target nucleic acid from the sample without extracting the target nucleic acid from the sample;
- (b) thermocycling the sample from step (a) such that the target nucleic acid, if present, is amplified;
- and (c) detecting the amplified target nucleic acid, if present, produced from step (b), wherein nucleic acid in the sample is not extracted from the sample prior to amplification, and wherein the buffer comprises at least one of component selected from the group consisting of KCl, bovine serum albumin and a cationic surfactant.
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Dependencies are inferred from the claim text.