Sulfur atom modification on thymine improves the specificity and sensitivity of DNA polymerization and detection

Area of Science:

• Molecular Biology
• Biochemistry
• Genetics

Background:

• Accurate base-pairing is crucial for molecular recognition, DNA polymerization, and amplification.
• Mismatched base pairs, like the T/G wobble pair, can lead to non-specific interactions.
• Existing methods face limitations in sensitivity and specificity due to these mismatches.

Purpose of the Study:

• To investigate the potential of 2-S-TTP (S-TTP) in minimizing T/G mismatches during DNA polymerization.
• To evaluate the impact of S-TTP on DNA polymerization specificity and detection sensitivity.
• To compare the performance of S-TTP with canonical TTP in nucleic acid detection.

Main Methods:

• Utilizing 2-S-TTP (S-TTP) in DNA polymerization reactions.
• Assessing DNA polymerization specificity and efficiency.
• Evaluating detection sensitivity for pathogens, including COVID-19, using S-TTP.

Main Results:

• S-TTP significantly minimizes T/G mismatches, enhancing DNA polymerization specificity.
• Detection sensitivity is improved up to 20-fold without compromising polymerization efficiency.
• The S-TTP strategy enables pathogen detection at single-digit copy numbers (up to 5 copies).

Conclusions:

• S-TTP offers substantially higher specificity and sensitivity in DNA polymerization and nucleic acid detection compared to canonical TTP.
• This approach overcomes limitations of conventional RT-PCR for detecting low-copy number pathogens.
• S-TTP represents a promising advancement for sensitive and specific molecular diagnostics.