We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Introducing Triplex Forming Oligonucleotide into Loop-Mediated Isothermal Amplification for Developing a Lateral Flow Biosensor for Streptococci Detection.
- Authors
Chang, Wei; Chou, Po-Hao; Wu, Cai-Tong; Song, Jheng-Da; Tsai, Kun-Nan; Chiou, Chiuan-Chian
- Abstract
Loop-mediated isothermal amplification (LAMP) technology is extensively utilized for the detection of infectious diseases owing to its rapid processing and high sensitivity. Nevertheless, conventional LAMP signaling methods frequently suffer from a lack of sequence specificity. This study integrates a triplex-forming oligonucleotide (TFO) probe into the LAMP process to enhance sequence specificity. This TFO-LAMP technique was applied for the detection of Group B Streptococcus (GBS). The TFO probe is designed to recognize a specific DNA sequence, termed the TFO targeting sequence (TTS), within the amplified product, facilitating detection via fluorescent instrumentation or lateral flow biosensors. A screening method was developed to identify TFO sequences with high affinity to integrate TFO into LAMP, subsequently incorporating a selected TTS into an LAMP primer. In the TFO-LAMP assay, a FAM-labeled TFO is added to target the TTS. This TFO can be captured by an anti-FAM antibody on lateral flow test strips, thus creating a nucleic acid testing biosensor. The efficacy of the TFO-LAMP assay was confirmed through experiments with specimens spiked with varying concentrations of GBS, demonstrating 85% sensitivity at 300 copies and 100% sensitivity at 30,000 copies. In conclusion, this study has successfully developed a TFO-LAMP technology that offers applicability in lateral flow biosensors and potentially other biosensor platforms.
- Subjects
STREPTOCOCCUS agalactiae; OLIGONUCLEOTIDES; STREPTOCOCCUS; BIOSENSORS; POLYMERASES; NUCLEOTIDE sequence; NUCLEIC acids
- Publication
Biosensors (2079-6374), 2024, Vol 14, Issue 5, p257
- ISSN
2079-6374
- Publication type
Article
- DOI
10.3390/bios14050257