Rapid diagnostic electrochemical biosensing targeted with antisense oligonucleotides
AssigneesUniversity of Maryland, Baltimore, University of Maryland, Baltimore County
InventorsDipanjan PAN, Maha Mohammad Shehadeh ALAFEEF, Ketan DIGHE, Parikshit MOITRA
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Abstract
The present disclosure relates to electrochemical biosensing systems and methods that can be adapted to accurately and rapidly detect a target gene in clinical samples, using anti-sense oligonucleotides for selective detection of biological pathogens.
Claims20
An electrochemical biosensor for use in the detection of a biological pathogen in a sample, the electrochemical biosensor comprising:
- a) a sensing element comprising a plurality of first anti-sense oligonucleotides, the sequence of which is complementary to a first nucleic acid sequence in a target gene of the biological pathogen;
- b) a first electrode connected to a first end of each of the plurality of anti-sense oligonucleotides;
- and
- c) a second electrode electrically connected to the first electrode;
- wherein contact of the sample with the first electrode causes binding of the plurality of first anti-sense oligonucleotides to the first nucleic acid sequence in the target gene, to provide a signal to identify presence of the biological pathogen.
A method of detecting a biological pathogen in a sample, the method comprising:
- a) providing a plurality of first anti-sense oligonucleotides having a sequence complementary to a first nucleic acid sequence in a target gene of the biological pathogen;
- b) providing a first electrode and a second electrode electrically connected to one another;
- c) connecting a first end of the pluralities of the first anti-sense oligonucleotides to the first electrode;
- and
- d) contacting the first electrode to the sample;
- e) measuring a signal from the first and second electrodes;
- wherein the binding of the target gene of the biological pathogen to the plurality of first anti-sense oligonucleotides provides the signal identifying the presence of the biological pathogen in the sample.
A method of selecting at least one anti-sense oligonucleotide probe for use in detection of a biological pathogen, comprising:
- a) identifying a target gene in the biological pathogen;
- b) obtaining the nucleic acid sequence of the target gene;
- c) producing a library of anti-sense oligonucleotides of a length of about 20 nucleotides, wherein the sequence of each anti-sense oligonucleotide is complementary to a section of the nucleic acid sequence in the target gene and wherein:
- i. guanine and cysteine form from 40 to 60 percent of each anti-sense oligonucleotide in the library;
- ii. none of the anti-sense oligonucleotides in the library are complementary to a section of the target gene with the sequence GGGG;
- iii. the average unpaired probability of each of the anti-sense oligonucleotides in the library is at least 0.5;
- d) ranking the anti-sense oligonucleotides in the library in descending order of average unpaired probability;
- and
- e) selecting the at least one anti-sense oligonucleotide probe from the anti-sense oligonucleotides in the library, wherein the binding energy of the at least one anti-sense oligonucleotide probe is less than −8 kcal/mol, compared with the nucleic acid sequence of the target gene.
Dependencies are inferred from the claim text.