US 12,435,356 B2
Target reporter constructs and uses thereof
Michael L. Metzker, Houston, TX (US); and Christopher August Weier, Houston, TX (US)
Assigned to RedVault Biosciences, LP, Houston, TX (US)
Filed by RedVault Biosciences, LP, Houston, TX (US)
Filed on Mar. 23, 2022, as Appl. No. 17/702,152.
Application 17/702,152 is a continuation of application No. 15/614,958, filed on Jun. 6, 2017, granted, now 11,299,769.
Claims priority of provisional application 62/346,334, filed on Jun. 6, 2016.
Prior Publication US 2022/0307076 A1, Sep. 29, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12Q 1/68 (2018.01); C12Q 1/6806 (2018.01); C12Q 1/6809 (2018.01); C12Q 1/6811 (2018.01); C12Q 1/6837 (2018.01); C12Q 1/6844 (2018.01); C12Q 1/6853 (2018.01); C12Q 1/686 (2018.01); C12Q 1/70 (2006.01); C12Q 1/6869 (2018.01); C12Q 1/6876 (2018.01)
CPC C12Q 1/6811 (2013.01) [C12Q 1/6806 (2013.01); C12Q 1/6809 (2013.01); C12Q 1/6837 (2013.01); C12Q 1/6844 (2013.01); C12Q 1/6853 (2013.01); C12Q 1/686 (2013.01); C12Q 1/703 (2013.01); C12Q 1/6869 (2013.01); C12Q 1/6876 (2013.01); C12Q 2600/178 (2013.01)] 23 Claims
 
1. A method for determining the presence or the absence of a pathogenic nucleic acid in a sample comprising:
(a) obtaining a target reporter construct (TRC), the TRC containing
(i) a target sequence complementary to the pathogenic nucleic acid;
(ii) a bridge sequence forming double-stranded portion of the TRC; and
(iii) an access sequence comprising a multifunctional probe sequence, wherein the bridge sequence comprises a palindromic sequence for the formation of nanospheres during replication;
(b) contacting the target reporter construct (TRC) with a sample to form a hybridized TRC if a pathogenic nucleic acid is present in the sample;
(c) performing rolling circle replication (RCR) on the hybridized TRC, thereby obtaining a replicated TRC if a pathogenic nucleic acid is present in the sample, the pathogenic nucleic acid serving as a primer for the RCR;
(d) contacting the replicated TRC with a detectable moiety, wherein the detectable moiety binds a probe sequence of the replicated TRC to form a bound detectable moiety if a pathogenic nucleic acid is present in the sample; and
(e) detecting for the bound detectable moiety, whereby detection of the bound detectable moiety corresponds to the presence of the pathogenic nucleic acid in the sample and the failure to detect the bound detectable moiety corresponds to the absence of the pathogenic nucleic acid in the sample.