Method of rapid extraction of DNA from saliva and qPCR amplification of group a Streptococcus DNA
AssigneeSalivIQ Diagnostics, Inc.
InventorJulia A. Halterman
Life of this patent
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Abstract
A method for processing a saliva sample for downstream DNA analysis. The method includes mixing the saliva sample with a chelating resin solution to form a mixture, incubating the mixture at a temperature of 120° C. or lower and duration of at least 1 minute, wherein the temperature and duration are inversely related, shaking the mixture before and after incubation for a period ranging from 0 seconds to 2 hours, separating the mixture into a supernatant and a sedimented chelating resin using either centrifugation or by allowing natural gravitational settling, and recovering the supernatant for subsequent analysis.
Claims20
A method for processing a saliva sample for subsequent bacterial DNA analysis, the method comprising:
- mixing the saliva sample with a chelating resin solution to form a mixture;
- incubating the mixture at a temperature of from 100° C. to 120° C. for a duration of from 30 seconds to 10 minutes, wherein the temperature and duration are inversely related;
- optionally shaking the mixture before and/or after incubation for a period up to 2 hours;
- separating the mixture into a supernatant and a sedimented chelating resin;
- and
- recovering the supernatant for subsequent bacterial DNA analysis.
A method for detecting bacterial DNA in a saliva sample comprising:
- mixing the saliva sample with a chelating resin solution to form a mixture;
- incubating the mixture at a temperature of from 100° C. to 120° C. for a duration of from 30 seconds to 10 minutes, wherein the temperature and duration are inversely related;
- separating the mixture into a supernatant and a sedimented chelating resin;
- and
- subjecting the supernatant to quantitative real-time polymerase chain reaction (qPCR) analysis to determine whether bacterial DNA is present in the supernatant,
- wherein incubating the mixture inactivates sufficient proteins present in the mixture such that qPCR analysis of the supernatant is not inhibited.
Dependencies are inferred from the claim text.