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

• Published in: Molecules (Basel, Switzerland) Switzerland), 2022-10, Vol.27 (20), p.7036
• Main Authors: Ma, Chun-Ho (Charlie), Li, Liejun, Cai, Shuheng, Lin, Pei, Lam, Wing-Ki (Kristy), Lee, Tsz-Him (Ronald), Kwok, Tsz-Kin (Ryan), Xie, Longxu, Kun, Tit-Sang (Tom), Tang, Ben-Zhong
• Format: Article
• Language: English
• Subjects: Accuracy; aggregation-induced emission; Alkaline phosphatase; Anemia; Aqueous solutions; Cervical cancer; Cervix; Colorimetry; Commercialization; Deoxyribonucleic acid; Diagnosis; Diagnostic systems; Disease; DNA; Emissions; Enzymes; flow-through hybridization; Fluorescence; Fluorescent dyes; Fluorescent indicators; Gene flow; Gene mutations; Genes; genetic diseases; Genomes; Genotype; Genotype & phenotype; Genotypes; Health risks; human immunodeficiency virus; Human papillomavirus; Hybridization; Infections; Medical research; Medical screening; Medicine, Experimental; membrane reader; Membranes; Methods; Molecular diagnostic techniques; Mutation; Nucleic acid hybridization; Papillomavirus infections; Phosphatase; Risk factors; Sickle cell disease
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules27207036

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.