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US 11,713,330 B2Granted
Methods and devices for ultrasensitive direct detection of microorganisms
AssigneeHelixBind, Inc.
InventorsAlon Singer, David Steinmiller, Nadish Goyal, Jork Nolling
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Abstract
The present disclosure generally relates to the field of ultrasensitive microbial pathogen detection and identification utilizing genomic sequence recognition.
Claims20
An ultrasensitive method of detecting one or more species of microbial cells in a biological sample comprising mammalian cells, the method comprising:
- selectively lysing the mammalian cells in the biological sample, including cells which comprise eukaryotic DNA, by contacting the biological sample with a composition;
- amplifying a plurality of microbial genetic materials in the biological sample;
- and
- detecting the amplified microbial genetic material,
- wherein the composition consists essentially of:
- a magnesium salt;
- a buffering agent;
- water;
- and
- a compound of Formula 1:
- wherein R1 is selected from the group consisting of optionally substituted, branched or unbranched, saturated or unsaturated C1-C8 aliphatic;
- optionally substituted, saturated or unsaturated C3-C14 carbocyclic;
- optionally substituted, saturated or unsaturated 3-8 membered heterocyclic;
- optionally substituted, branched or unbranched, saturated or unsaturated ((Ra)q—(C═O)—(Ra)q)p;
- optionally substituted C6-C14 aryl;
- and optionally substituted 3-8 membered heteroaryl;
- and/or any suitable combinations thereof;
- wherein R2 is selected from the group consisting of hydrogen;
- optionally substituted, branched or unbranched, saturated or unsaturated C1-C28 aliphatic;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb)n—O—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb)n—NH—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb—O)n—S—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(S—Rb)n—S—Rb)p;
- optionally substituted C6-C14 aryl;
- optionally substituted 3-8 membered heteroaryl;
- optionally substituted, saturated or unsaturated C3-C14 carbocyclic;
- optionally substituted, saturated or unsaturated 3-8 membered heterocyclic;
- optionally substituted, branched or unbranched, saturated or unsaturated —(C═O)—(Rb);
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—O—(Ra)q)p—;
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—NH—(Ra)q)p—;
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—N(Ra)—(Ra)q)p—;
- and optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—S—(Ra)q)p—;
- and/or any suitable combinations thereof;
- wherein each occurrence of Ra is independently C1-C8 aliphatic or C6-C14 aryl;
- wherein each occurrence of Rb is independently C1-C15 aliphatic or C6-C14 aryl;
- wherein each occurrence of subscript q is independently an integer between 0 and 1, inclusive;
- wherein each occurrence of subscript p is independently an integer between 1 and 6, inclusive;
- and
- wherein each occurrence of subscript n is independently an integer between 0 and 14, inclusive.
36 more elements
A method of selectively lysing mammalians cells in a biological sample comprising mammalian cells comprising eukaryotic DNA and Borrelia cells, the method comprising contacting the biological sample with a composition, wherein the composition consists essentially of:
- a magnesium salt;
- a buffering agent;
- water;
- and
- a compound of Formula 1:
- wherein R1 is selected from the group consisting of optionally substituted, branched or unbranched, saturated or unsaturated C1-C8 aliphatic;
- optionally substituted, saturated or unsaturated C3-C14 carbocyclic;
- optionally substituted, saturated or unsaturated 3-8 membered heterocyclic;
- optionally substituted, branched or unbranched, saturated or unsaturated ((Ra)q—(C═O)—(Ra)q)p;
- optionally substituted C6-C14 aryl;
- and optionally substituted 3-8 membered heteroaryl;
- and/or any suitable combinations thereof;
- wherein R2 is selected from the group consisting of hydrogen;
- optionally substituted, branched or unbranched, saturated or unsaturated C1-C28 aliphatic;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb)n—O—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb)n—NH—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(O—Rb—O)n—S—Rb)p;
- optionally substituted, branched or unbranched, saturated or unsaturated —(Rb—(S—Rb)n—S—Rb)p;
- optionally substituted C6-C14 aryl;
- optionally substituted 3-8 membered heteroaryl;
- optionally substituted, saturated or unsaturated C3-C14 carbocyclic;
- optionally substituted, saturated or unsaturated 3-8 membered heterocyclic;
- optionally substituted, branched or unbranched, saturated or unsaturated —(C═O)—(Rb);
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—O—(Ra)q)p—;
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—NH—(Ra)q)p—;
- optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—N(Ra)—(Ra)q)p—;
- and optionally substituted, branched or unbranched, saturated or unsaturated —((Ra)q—S—(Ra)q)p—;
- and/or any suitable combinations thereof;
- wherein each occurrence of Ra is independently C1-C8 aliphatic or C6-C14 aryl;
- wherein each occurrence of Rb is independently C1-C15 aliphatic or C6-C14 aryl;
- wherein each occurrence of subscript q is independently an integer between 0 and 1, inclusive;
- wherein each occurrence of subscript p is independently an integer between 1 and 6, inclusive;
- and
- wherein each occurrence of subscript n is independently an integer between 0 and 14, inclusive.
31 more elements
Dependencies are inferred from the claim text.