Polynucleotides for amplification and detection of human beta actin
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 primers comprising set-29, wherein set-29 comprises primer sequences of SEQ ID NO: 132 and SEQ ID NO: 133.
2. The composition of claim 1 , further comprising a probe.
3. The composition at claim 1 , further comprising a labeled probe, wherein the probe sequence comprises SEQ ID NO:184.
4. The composition of claim 2 , wherein the probe is a molecular beacon comprising a fluorophore and a quencher.
5. A molecular beacon probe comprising a fluorophore and a quencher, wherein the probe sequence comprises SEQ ID NO: 184.
6. A kit comprising a composition according to claim 1 .
7. 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 isothermal loop mediated amplification (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,
wherein said sequence-specific LAMP primer set is set-29, wherein set-29 comprises primer sequences of SEQ ID NO: 132 and SEQ ID NO: 133.
8. The method of claim 7 , wherein detecting the presence or absence of the amplification product comprises hybridizing the amplified product with a molecular beacon probe, wherein the probe sequence comprises SEQ ID NO: 184.