Development of an HPV Genotype Detection Platform Based on Aggregation-Induced Emission (AIE) and Flow-Through Hybridization Technologies

Abstract

Genetic mutations can cause life-threatening diseases such as cancers and sickle cell anemia. Gene detection is thus of importance for disease-risk prediction or early diagnosis and treatment. Apart from genetic defects, gene detection techniques can also be applied to gene-related diseases with high risk to human health such as human papillomavirus (HPV) infection. HPV infection has been strongly linked to cervical cancer. To achieve a high-throughput HPV gene detection platform, the flow-through hybridization system appears to be one of the commercialized diagnostic techniques for this purpose. The flow-through hybridization technique is based on a vacuum-guided flow of DNA fragments which is continuously directed toward the oligoprobes that are immobilized on the testing membrane. However, the conventional colorimetric method and signal read-out approach suffers a problem of low sensitivity. On the contrary, fluorescence approaches allow more sensitive detection and broad sensing ranges. In this work, a fluorescent dye HCAP, which possesses aggregation-induced emission (AIE) properties and is responsive to alkaline phosphatase, was developed and applied to the flow-through hybridization platform to achieve HPV genome diagnosis of clinical samples. Also, an automatic membrane reader was constructed based on the AIE-based diagnosis platform which can identify the diagnostic result of patient DNA with a total concordance rate of 100% in the clinical trial.

Development and validation of real-time recombinase polymerase amplification-based assays for detecting HPV16 and HPV18 DNA

“…In clinical and field conditions, there is a demand for more convenient rapid point-of-care testing (POCT), with simpler equipment requirements, and lower costs. Novel HPV detection methods, such as droplet digital PCR (ddPCR) ( 12 ), AuNP (gold nanoparticle)-HPV probe characterization assays ( 13 ), aggregation-induced emission (AIE) combined with flow-through hybridization technologies ( 14 ), recombinase polymerase amplification (RPA)-based assays ( 15 ), loop-mediated amplification (LAMP) reactions ( 16 ), and RPA combined with CRISPR-Cas12 ( 17 ), have been developed, with the aim of shortening the time required to obtain results, simplifying the interpretation of results and reducing diagnostic costs. Until now, a total of 15 articles have applied RPA to HPV detection ( Table 1 ).…”