Polynucleotides for the amplification and detection of human beta actin

Abstract

Disclosed herein are primers and probes related to the detection of beta actin [Homo sapiens (human)] via nucleic acid amplification testing (NAAT), for example to amplify and determine the presence of β-actin present in test samples. Specifically, the present disclosure describes primers and probes that bind to the beta actin gene for detection via loop mediated isothermal amplification (LAMP) and molecular beacon hybridization.

1 . A composition comprising a set of polynucleotides selected from the group consisting of Set-1 through Set-29. 
     
     
         2 . The composition of  claim 1 , further comprising a probe. 
     
     
         3 . The composition of  claim 2 , wherein the probe comprises a label. 
     
     
         4 . The composition of  claim 3 , wherein the probe is a labeled polynucleotide. 
     
     
         5 . The composition of  claim 4 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135, nucleotides 6-30 of SEQ ID NO: 136, nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 143, nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, nucleotides 8-30 of SEQ ID NO: 146, 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-32 of SEQ ID NO: 151, nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 7-25 of SEQ ID NO: 159, nucleotides 8-22 of SEQ ID NO: 160, nucleotides 6-22 of SEQ ID NO: 161, nucleotides 3-22 of SEQ ID NO: 162, nucleotides 8-28 of SEQ ID NO: 163, nucleotides 3-28 of SEQ ID NO: 164, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 3-22 of SEQ ID NO: 170, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 4-23 of SEQ ID NO: 177, nucleotides 4-34 of SEQ ID NO: 178, nucleotides 3-27 of SEQ ID NO: 179, nucleotides 2-27 of SEQ ID NO: 180, nucleotides 5-33 of SEQ ID NO: 181, nucleotides 3-30 of SEQ ID NO: 182, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184. 
     
     
         6 . The composition of  claim 4 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         7 . The composition of  claim 4 , wherein the label is a fluorophore. 
     
     
         8 . The composition of  claim 7 , wherein the fluorophore is covalently attached to a terminus of the polynucleotide. 
     
     
         9 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184, and wherein the set of polynucleotides is selected from the group consisting of Sets 9-12, Set-17, and Sets 22-29. 
     
     
         10 . The composition of  claim 9 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 183, and SEQ ID NO: 184. 
     
     
         11 . The composition of  claim 10 , wherein the sequence of the labeled polynucleotides is SEQ ID NO: 184, and the set of polynucleotides is Set-29. 
     
     
         12 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, and nucleotides 1-23 of SEQ ID NO: 154, and wherein the set of polynucleotides is selected from the group consisting of Sets 9-11, Set-17, and Sets 23-29. 
     
     
         13 . The composition of  claim 12 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 152, SEQ ID NO: 153, and SEQ ID NO: 154. 
     
     
         14 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, and nucleotides 8-30 of SEQ ID NO: 146, and wherein the set of polynucleotides is selected from the group consisting of Set-4, Sets 9-12, Set-17, and Sets 22-26. 
     
     
         15 . The composition of  claim 14 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 144, SEQ ID NO: 145, and SEQ ID NO: 146. 
     
     
         16 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising nucleotides 3-22 of SEQ ID NO: 170, and wherein the set of polynucleotides is selected from the group consisting of Set-5, Set-12, Set-17, and Sets 22-25. 
     
     
         17 . The composition of  claim 16 , wherein the labeled polynucleotide comprises SEQ ID NO: 170. 
     
     
         18 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 2-27 of SEQ ID NO: 180 and nucleotides 3-30 of SEQ ID NO: 182, and wherein the set of polynucleotides is selected from the group consisting of Sets 6-8, Set-15, and Set-16. 
     
     
         19 . The composition of  claim 18 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 180 and SEQ ID NO: 182. 
     
     
         20 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 4-34 of SEQ ID NO: 178 and nucleotides 5-33 of SEQ ID NO: 181, and wherein the set of polynucleotides is selected from the group consisting of Sets 6-11 and Sets 28-29. 
     
     
         21 . The composition of  claim 20 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 178 and SEQ ID NO: 181. 
     
     
         22 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising nucleotides 3-27 of SEQ ID NO: 179, and wherein the set of polynucleotides is selected from the group consisting of Sets 6-9 and Set-11. 
     
     
         23 . The composition of  claim 22 , wherein the labeled polynucleotide comprises SEQ ID NO: 179. 
     
     
         24 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising nucleotides 4-23 of SEQ ID NO: 177, and wherein the set of polynucleotides is selected from the group consisting of Sets 6-9, Set-11, and Set-28. 
     
     
         25 . The composition of  claim 24 , wherein the labeled polynucleotide comprises SEQ ID NO: 177. 
     
     
         26 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 7-25 of SEQ ID NO: 159 and nucleotides 8-22 of SEQ ID NO: 160, and wherein the set of polynucleotides is selected from the group consisting of Sets 13-15. 
     
     
         27 . The composition of  claim 26 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 159 and SEQ ID NO: 160. 
     
     
         28 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 6-22 of SEQ ID NO: 161, and nucleotides 3-22 of SEQ ID NO: 162, and wherein the set of polynucleotides is selected from the group consisting of Sets 13-16. 
     
     
         29 . The composition of  claim 28 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 161, and SEQ ID NO: 162. 
     
     
         30 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group of nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 151, and nucleotides 4-24 of SEQ ID NO: 153, and wherein the set of polynucleotides is Set-3. 
     
     
         31 . The composition of  claim 30 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 151, and SEQ ID NO: 153. 
     
     
         32 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 8-28 of SEQ ID NO: 163 and nucleotides 3-28 of SEQ ID NO: 164, and wherein the set of polynucleotides is selected from the group consisting of Set-1, Set-2, and Sets 18-20. 
     
     
         33 . The composition of  claim 32 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 163 and SEQ ID NO: 164. 
     
     
         34 . The composition of  claim 1 , further comprising a labeled polynucleotide comprising a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135 and nucleotides 6-30 of SEQ ID NO: 136, and wherein the set of polynucleotides is selected from the group consisting of Set-1, Set-2 and Sets 18-21. 
     
     
         35 . The composition of  claim 34 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 135 and SEQ ID NO: 136. 
     
     
         36 . The composition of  claim 2 , wherein the probe is a molecular beacon comprising a fluorophore, a quencher, and a polynucleotide. 
     
     
         37 . The composition of  claim 36 , wherein the molecular beacon comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135, nucleotides 6-30 of SEQ ID NO: 136, nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 143, nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, nucleotides 8-30 of SEQ ID NO: 146, 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-32 of SEQ ID NO: 151, nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 7-25 of SEQ ID NO: 159, nucleotides 8-22 of SEQ ID NO: 160, nucleotides 6-22 of SEQ ID NO: 161, nucleotides 3-22 of SEQ ID NO: 162, nucleotides 8-28 of SEQ ID NO: 163, nucleotides 3-28 of SEQ ID NO: 164, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 3-22 of SEQ ID NO: 170, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 4-23 of SEQ ID NO: 177, nucleotides 4-34 of SEQ ID NO: 178, nucleotides 3-27 of SEQ ID NO: 179, nucleotides 2-27 of SEQ ID NO: 180, nucleotides 5-33 of SEQ ID NO: 181, nucleotides 3-30 of SEQ ID NO: 182, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184. 
     
     
         38 . The composition of  claim 37 , wherein the molecular beacon comprises a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         39 . The composition of  claim 38 , wherein the polynucleotide sequence consists of SEQ ID NO: 184. 
     
     
         40 . A molecular beacon comprising a fluorophore, a quencher, and a polynucleotide, wherein the polynucleotide comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135, nucleotides 6-30 of SEQ ID NO: 136, nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 143, nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, nucleotides 8-30 of SEQ ID NO: 146, 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-32 of SEQ ID NO: 151, nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 7-25 of SEQ ID NO: 159, nucleotides 8-22 of SEQ ID NO: 160, nucleotides 6-22 of SEQ ID NO: 161, nucleotides 3-22 of SEQ ID NO: 162, nucleotides 8-28 of SEQ ID NO: 163, nucleotides 3-28 of SEQ ID NO: 164, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 3-22 of SEQ ID NO: 170, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 4-23 of SEQ ID NO: 177, nucleotides 4-34 of SEQ ID NO: 178, nucleotides 3-27 of SEQ ID NO: 179, nucleotides 2-27 of SEQ ID NO: 180, nucleotides 5-33 of SEQ ID NO: 181, nucleotides 3-30 of SEQ ID NO: 182, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184. 
     
     
         41 . The molecular beacon of  claim 40 , wherein the polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         42 . The molecular beacon of  claim 41 , wherein the polynucleotide consists a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         43 . The molecular beacon of  claim 40 , wherein the fluorophore is FAM and the quencher is BHQ1. 
     
     
         44 . The molecular beacon of  claim 40 , wherein the fluorophore is ATTO 565 or Alexa 594 and the quencher is BHQ1 or BHQ2. 
     
     
         45 . A method of detecting β-actin in a test sample, the method comprising:
 (a) extracting nucleic acid from the test sample; 
 (b) amplifying a target sequence by reacting the nucleic acid extracted in step (a) with a reaction mixture comprising a strand displacement DNA polymerase and a sequence specific primer set, wherein said sequence-specific primer set is selected from the group consisting of Set-1 through Set-29; and 
 (c) detecting the presence or absence of an amplified product of step (b); wherein the presence of said amplification product is indicative of the presence of β-actin in the test sample. 
 
     
     
         46 . The method of  claim 45 , wherein the amplification in step (b) of the target sequence is performed between about 60° C. and about 67° C. for less than 30 minutes. 
     
     
         47 . The method of  claim 46 , wherein the amplification step is performed for less than fifteen minutes. 
     
     
         48 . The method of  claim 47 , wherein the amplification step is performed for less than twelve minutes. 
     
     
         49 . The method of  claim 48 , wherein the amplification step is performed for less than nine minutes. 
     
     
         50 . The method of  claim 45 , wherein the reaction mixture further comprises a reverse transcriptase. 
     
     
         51 . The method of  claim 45 , wherein detecting the presence or absence of the amplification product comprises hybridizing the amplified product with a probe comprising a polynucleotide attached to a label. 
     
     
         52 . The method of  claim 51 , wherein the label is a fluorophore. 
     
     
         53 . The method of  claim 52 , wherein the fluorophore is covalently attached to a terminus of the polynucleotide. 
     
     
         54 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135, nucleotides 6-30 of SEQ ID NO: 136, nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 143, nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, nucleotides 8-30 of SEQ ID NO: 146, 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-32 of SEQ ID NO: 151, nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 7-25 of SEQ ID NO: 159, nucleotides 8-22 of SEQ ID NO: 160, nucleotides 6-22 of SEQ ID NO: 161, nucleotides 3-22 of SEQ ID NO: 162, nucleotides 8-28 of SEQ ID NO: 163, nucleotides 3-28 of SEQ ID NO: 164, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 3-22 of SEQ ID NO: 170, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 4-23 of SEQ ID NO: 177, nucleotides 4-34 of SEQ ID NO: 178, nucleotides 3-27 of SEQ ID NO: 179, nucleotides 2-27 of SEQ ID NO: 180, nucleotides 5-33 of SEQ ID NO: 181, nucleotides 3-30 of SEQ ID NO: 182, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184. 
     
     
         55 . The method of  claim 54 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         56 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184, and wherein the sequence specific primer set is selected from the group consisting of Sets 9-12, Set-17, and Sets 22-29. 
     
     
         57 . The method of  claim 56 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 183, SEQ ID NO: 184. 
     
     
         58 . The method of  claim 57 , wherein the sequence of the labeled polynucleotide is SEQ ID NO: 184, and the sequence-specific primer set is Set-29. 
     
     
         59 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, and wherein the sequence-specific primer set is selected from the group consisting of Sets 9-11, Set-17, and Sets 23-29. 
     
     
         60 . The method of  claim 59 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 152, SEQ ID NO: 153, and SEQ ID NO: 154. 
     
     
         61 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, and nucleotides 8-30 of SEQ ID NO: 146, and wherein the sequence-specific primer set is selected from the group consisting of Set-4, Sets 9-12, Set-17, and Sets 22-26. 
     
     
         62 . The method of  claim 61 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 144, SEQ ID NO: 145, and SEQ ID NO: 146. 
     
     
         63 . The method of  claim 51 , wherein the labeled polynucleotide comprises nucleotides 3-22 of SEQ ID NO: 170, and wherein the sequence-specific primer set is selected from the group consisting of Set-5, Set-12, Set-17, and Sets 22-25. 
     
     
         64 . The method of  claim 63 , wherein the labeled polynucleotide comprises SEQ ID NO: 170. 
     
     
         65 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 2-27 of SEQ ID NO: 180 and nucleotides 3-30 of SEQ ID NO: 182, and wherein the sequence-specific set is selected from the group consisting of Sets 6-8, Set-15, and Set-16. 
     
     
         66 . The method of  claim 65 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 180 and SEQ ID NO: 182. 
     
     
         67 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 4-34 of SEQ ID NO: 178 and nucleotides 5-33 of SEQ ID NO: 181, and wherein the sequence-specific primer set is selected from the group consisting of Sets 6-11 and Sets 28-29. 
     
     
         68 . The method of  claim 67 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 178 and SEQ ID NO: 181. 
     
     
         69 . The method of  claim 51 , wherein the labeled polynucleotide comprises nucleotides 3-27 of SEQ ID NO: 179, and wherein the sequence-specific primer set is selected from the group consisting of Sets 6-9 and Set-11. 
     
     
         70 . The method of  claim 69 , wherein the labeled polynucleotide comprises SEQ ID NO: 179. 
     
     
         71 . The method of  claim 51 , wherein the labeled polynucleotide comprises nucleotides 4-23 of SEQ ID NO: 177, and wherein the sequence-specific primer set is selected from the group consisting of Sets 6-9, Set-11, and Set-28. 
     
     
         72 . The method of  claim 71 , wherein the labeled polynucleotide comprises SEQ ID NO: 177. 
     
     
         73 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 7-25 of SEQ ID NO: 159 and nucleotides 8-22 of SEQ ID NO: 160, and wherein the sequence-specific primer set is selected from the group consisting of Sets 13-15. 
     
     
         74 . The method of  claim 73 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 159 and SEQ ID NO: 160. 
     
     
         75 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 6-22 of SEQ ID NO: 161, and nucleotides 3-22 of SEQ ID NO: 162, and wherein the sequence-specific primer set is selected from the group consisting of Sets 13-16. 
     
     
         76 . The method of  claim 75 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 161, and SEQ ID NO: 162. 
     
     
         77 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 151, and nucleotides 4-24 of SEQ ID NO: 153, and wherein the sequence-specific primer set is Set-3. 
     
     
         78 . The method of  claim 77 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 137, SEQ ID NO: 138, SEQ ID NO: 139, SEQ ID NO: 140, SEQ ID NO: 141, SEQ ID NO: 142, SEQ ID NO: 151, and SEQ ID NO: 153. 
     
     
         79 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 8-28 of SEQ ID NO: 163 and nucleotides 3-28 of SEQ ID NO: 164, and wherein the sequence-specific primer set is selected from the group consisting of Set-1, Set-2, and Sets 18-20. 
     
     
         80 . The method of  claim 79 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 163 and SEQ ID NO: 164. 
     
     
         81 . The method of  claim 51 , wherein the labeled polynucleotide comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135 and nucleotides 6-30 of SEQ ID NO: 136, and wherein the sequence-specific primer set is selected from the group consisting of Sets 1-2 and Sets 18-21. 
     
     
         82 . The method of  claim 81 , wherein the labeled polynucleotide comprises a sequence from the group consisting of SEQ ID NO: 135 and SEQ ID NO: 136. 
     
     
         83 . A kit comprising a composition according to  claim 1 . 
     
     
         84 . The kit of  claim 83 , further comprising a strand displacement polymerase. 
     
     
         85 . The kit of  claim 84 , further comprising a molecular beacon comprising a fluorophore, a quencher, and a polynucleotide, wherein the polynucleotide comprises a sequence selected from the group consisting of nucleotides 6-33 of SEQ ID NO: 135, nucleotides 6-30 of SEQ ID NO: 136, nucleotides 7-27 of SEQ ID NO: 137, nucleotides 7-26 of SEQ ID NO: 138, nucleotides 1-26 of SEQ ID NO: 139, nucleotides 7-26 of SEQ ID NO: 140, nucleotides 7-30 of SEQ ID NO: 141, nucleotides 6-30 of SEQ ID NO: 142, nucleotides 8-32 of SEQ ID NO: 143, nucleotides 8-30 of SEQ ID NO: 144, nucleotides 6-31 of SEQ ID NO: 145, nucleotides 8-30 of SEQ ID NO: 146, 8-22 of SEQ ID NO: 147, nucleotides 8-24 of SEQ ID NO: 148, nucleotides 7-27 of SEQ ID NO: 149, nucleotides 8-21 of SEQ ID NO: 150, nucleotides 8-32 of SEQ ID NO: 151, nucleotides 1-24 of SEQ ID NO: 152, nucleotides 4-24 of SEQ ID NO: 153, nucleotides 1-23 of SEQ ID NO: 154, nucleotides 8-26 of SEQ ID NO: 155, nucleotides 7-29 of SEQ ID NO: 156, nucleotides 8-28 of SEQ ID NO: 157, nucleotides 8-29 of SEQ ID NO: 158, nucleotides 7-25 of SEQ ID NO: 159, nucleotides 8-22 of SEQ ID NO: 160, nucleotides 6-22 of SEQ ID NO: 161, nucleotides 3-22 of SEQ ID NO: 162, nucleotides 8-28 of SEQ ID NO: 163, nucleotides 3-28 of SEQ ID NO: 164, nucleotides 5-25 of SEQ ID NO: 165, nucleotides 5-26 of SEQ ID NO: 166, nucleotides 5-20 of SEQ ID NO: 167, nucleotides 4-22 of SEQ ID NO: 168, nucleotides 7-22 of SEQ ID NO: 169, nucleotides 3-22 of SEQ ID NO: 170, nucleotides 7-28 of SEQ ID NO: 171, nucleotides 6-27 of SEQ ID NO: 172, nucleotides 7-29 of SEQ ID NO: 173, nucleotides 5-27 of SEQ ID NO: 174, nucleotides 7-29 of SEQ ID NO: 175, nucleotides 6-28 of SEQ ID NO: 176, nucleotides 4-23 of SEQ ID NO: 177, nucleotides 4-34 of SEQ ID NO: 178, nucleotides 3-27 of SEQ ID NO: 179, nucleotides 2-27 of SEQ ID NO: 180, nucleotides 5-33 of SEQ ID NO: 181, nucleotides 3-30 of SEQ ID NO: 182, nucleotides 9-34 of SEQ ID NO: 183, and nucleotides 8-28 of SEQ ID NO: 184. 
     
     
         86 . The kit of  claim 85 , wherein the polynucleotide comprises a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         87 . The kit of  claim 86 , wherein the polynucleotide consists of a sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         88 . The kit of  claim 87 , wherein the polynucleotide consists of SEQ ID NO: 184 and the set of polynucleotides is Set-29. 
     
     
         89 . A method of detecting β-actin in a test sample, the method comprising:
 (a) extracting nucleic acid from the test sample; 
 (b) amplifying a target sequence by reacting nucleic acid extracted in step (a) for less than ten minutes with a reaction mixture comprising a strand displacement DNA polymerase and a sequence specific LAMP primer set; and 
 (c) detecting the presence or absence of an amplified product of step (b); wherein the presence of said amplification product is indicative of the presence of β-actin in the test sample. 
 
     
     
         90 . The method of  claim 89 , wherein the amplifying step comprises reacting the nucleic acid extracted in step (a) with a reaction mixture comprising a strand displacement DNA polymerase and a sequence-specific primer set, wherein said sequence-specific primer set is selected from the group consisting of Set-1 through Set-29. 
     
     
         91 . The method of  claim 90 , wherein detecting the presence or absence of the amplification product comprises hybridizing the amplified product with a molecular beacon comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO: 135 through SEQ ID NO: 184. 
     
     
         92 . The method of  claim 90 , wherein detecting the presence or absence of the amplification product comprises hybridizing the amplified product with a molecular beacon comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO: 147, SEQ ID NO: 148, SEQ ID NO: 149, SEQ ID NO: 150, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 165, SEQ ID NO: 166, SEQ ID NO: 167, SEQ ID NO: 168, SEQ ID NO: 169, SEQ ID NO: 171, SEQ ID NO: 172, SEQ ID NO: 173, SEQ ID NO: 174, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 183, and SEQ ID NO: 184.