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Updated: Apr 30, 2026
The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform
Published on: September 27, 2016
Enhanced SfaTnpB enables single-base-specific, one-pot nucleic acid detection for high-sensitivity diagnostics
Bingrong Xu1,2, Sheng Li1, Yong Li2
1Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education & Key Laboratory of Swine Genetics and Breeding of Ministry of Agriculture and Rural Affairs, Huazhong Agricultural University, Wuhan 430070, P.R. China.
We developed a new molecular detection system, TOPIC, using SfaTnpB for highly specific nucleic acid detection. This system accurately identifies human papillomavirus subtypes and African swine fever virus, offering potential for point-of-care diagnostics.
Area of Science:
- Molecular Diagnostics
- Nucleic Acid Detection
- Biotechnology
Background:
- CRISPR/Cas12 systems offer nucleic acid detection but lack specificity and compatibility for clinical applications.
- Limitations include poor single-nucleotide specificity, limited subtype discrimination, and incompatibility with one-pot assays.
Purpose of the Study:
- To develop a novel nucleic acid detection system with enhanced specificity and compatibility.
- To engineer a highly active and specific SfaTnpB enzyme system for precise molecular diagnostics.
Main Methods:
- Engineered SfaTnpB (transposon-associated transposase B) and its guide RNA (ωRNA) to create an enhanced system (enSfaTnpB).
- Developed TOPIC (TnpB-based One-Pot nucleIC acid detection) by combining enSfaTnpB with amplification methods (RAA or LAMP).
- Validated TOPIC for detecting human papillomavirus (HPV) subtypes and African swine fever virus DNA.
Main Results:
- The enSfaTnpB system demonstrated robust single-base mismatch discrimination and broad temperature tolerance.
- TOPIC enabled ultrasensitive detection of HPV subtypes 16/18 (∼4 copies/μl) and African swine fever virus DNA (∼3 copies/μl).
- RAA-TOPIC accurately genotyped 14 high-risk HPV subtypes with high fidelity, showing concordance with qPCR.
Conclusions:
- TOPIC is a compact, programmable, and scalable molecular detection platform.
- The system shows significant potential for precision diagnostics and point-of-care testing, especially in resource-limited settings.
- SfaTnpB-based detection offers an advancement over CRISPR/Cas12 for specific nucleic acid identification.
