Shixian Liu,Tian Yu,Libing Song,Kourosh Kalantar-Zadeh,Guozhen Liu
Highly sensitive point-of-care early screening for high-risk human papillomavirus (HPV) infections is urgently needed, particularly in resource-limited settings. Nucleic acid amplification methods, especially CRISPR/Cas-based biosensors, have emerged as promising tools for sensitive HPV detection; however, current approaches typically rely on tedious tube-based formats coupled with lateral flow assays for signal readout in point-of-care testing (POCT). Here, we developed customized microfluidic paper-based analytical devices (μPADs) with valves that seamlessly integrated recombinase polymerase amplification (RPA) with CRISPR/Cas12a biosensing (RPA-CRISPR/Cas12a) on the filter paper substrate. This innovation achieved sensitive and cost-effective high-risk HPV detection in POCT. The RPA-CRISPR/Cas12a system with a linear reporter on μPADs, enabled fluorescence detection of the E7 gene, achieving a sensitivity of 1 pM at approximately 1 h. The sensitivity was further enhanced by introducing a circular reporter into the fluorescence-based RPA-CRISPR/Cas12a system on μPADs, enabling detection of the E7 gene with a detection limit of 1 fM and an assay time of 35 min. The system was validated using 50 cervical swab clinical samples, demonstrating 95% sensitivity and 100% specificity when compared to qPCR. This sample-to-answer detection platform holds significant promise for early screening of high-risk HPV infections in point-of-care scenarios.
Drug-induced phospholipidosis (PLD) is a storage disorder of lysosomes characterized by the excessive accumulation of phospholipids as a result of improper medical treatments. Although few evidences have supported that PLD can induce significant pathological consequences, this potential toxicity indeed can put off the drug discovery process. In this research, a high-throughput liposome electrokinetic chromatography (LEKC) method was validated to evaluate the PLD risk of drug candidates by screening drug-phospholipid interaction, which correlates to the phospholipidosis inducing risk. A statistical analysis based on the Spearman's correlation test showed that the retention factors (log k) of the tested drugs in the LEKC system and the literature reported in vivo and in vitro PLD data were highly correlated. In order to investigate the predictability of LEKC, the effect of liposome composition such as the molar ratio of phospholipids and the addition of cholesterol were also discussed in this study. The results indicated that the LEKC method could offer a fast, reliable and cost-effective screening tool for early prediction of the PLD inducing potential of drug candidates.
Beta-amyloid 42 (Aβ-42) peptide is an essential biomarker for Alzheimer's disease (AD) diagnosis. This study presents high sensitivity and stability sensors for Aβ-42 peptide detection, along with an integrated handheld analysis and display device. The sensors, with a wide response range and high accuracy, capable of measuring Aβ-42 peptide concentration at the femtogram level, could have enabled early screening for AD. Employing an in-situ grown three-dimensional sensor, the specific surface area of the sensor was increased by 200 % which, thereby, provided more antibody binding sites to enhance detection accuracy. This electrochemical sensor demonstrated a broad linear response range from 4.5 × 10 −8 pg/mL to 4.5 × 10 2 pg/mL and could accurately determine the concentration of spiked analytes (including artificial saliva, human plasma, and human blood). Furthermore, the sensor exhibited satisfactory stability, with only a 5 % sensitivity loss after 35 days of storage at 4 °C. Additionally, this handheld Aβ-42 peptide testing device incorporates a display for result delivery. Such innovative technology could fill the gap in the market for portable testing instruments and could provide an efficient and reliable solution for early AD screening.
危险品化学品经营许可证（带存储）