Intestinal flora plays a crucial role in the host’s intestinal health. Imbalances in the intestinal flora, when accompanied by inflammation, affect the host’s intestinal barrier function. Understanding it requires studying how living cells and tissues work in the context of living organs, but it is difficult to form the three-dimensional microstructure intestinal–vascular system by monolayer cell or co-culture cell models, and animal models are costly and slow. The use of microfluidic-based organ chips is a fast, simple, and high-throughput method that not only solves the affinity problem of animal models but the lack of microstructure problem of monolayer cells. In this study, we designed an embedded membrane chip to generate an in vitro gut-on-a-chip model. Human umbilical vein endothelial cells and Caco-2 were cultured in the upper and lower layers of the culture chambers in the microfluidic chip, respectively. The human peripheral blood mononuclear cells were infused into the capillary side at a constant rate using an external pump to simulate the in vitro immune system and the shear stress of blood in vivo. The model exhibited intestine morphology and function after only 5 days of culture, which is significantly less than the 21 days required for static culture in the Transwell® chamber. Furthermore, it was observed that drug-resistant bacteria triggered barrier function impairment and inflammation, resulting in enteritis, whereas probiotics (Lactobacillus rhamnosus GG) improved only partially. The use of Amikacin for enteritis is effective, whereas other antibiotic therapies do not work, which are consistent with clinical test results. This model may be used to explore intestinal ecology, host and intestinal flora interactions, and medication assessment.

Influenza is a zoonotic disease, infecting a wide variety of warm-blooded animals. It is caused by an influenza virus, which has been found with hundreds of subtypes. These subtypes are often associated with different sources of infection and possess complex courses of infection. In the early stage of influenza infection, rapid subtype detection is very practicable to prevent the disease from getting worse. Herein, we presented a high-throughput microfluidic centrifugal disc for rapid detection of influenza virus subtypes. The disc realized detection reagent preloads, automated reagent control, and RT-LAMP detections. Six kinds of highly pathogenic influenza viruses could be simultaneously identified, including influenza A subtypes H1, H3, H5, H7, and H9 and influenza B virus. Two different fluorescent dyes could be used on the disc for real-time detection or read by the naked eye. The performance of the disc was demonstrated by testing the clinical samples. The integrated centrifugal disc was expected for rapid detection of influenza virus subtypes to facilitate accurate drug usage in resource-constrained settings and contribute to reduce the risk of the influenza pandemic.

Integrated clustered regularly interspaced short palindromic repeat (CRISPR)–loop-mediated amplification (LAMP) technology is of great importance in CRISPR-based diagnostic systems, which urgently needs to be developed to improve diagnostic accuracy. A labor-free, contamination-free, and fully automated droplet manipulation platform for the CRISPR-LAMP technology has not been developed before. Herein, we propose a fully automated CRISPR-LAMP platform, which can precisely manipulate the CRISPR-LAMP droplet and perform combined reactions with high sensitivity and specificity. SARS-CoV-2 Spike T478K, D614G, P681R, and P681H mutations, typical point mutations of B.1.617.2 (Delta) and Omicron variants, are monitored with this platform with a detection limit of 102 copies/μL. Allele discrimination between the mutants and wild type is significant with the designed one/two-mismatch CRISPR RNA (crRNA) at a limit of 102 copies/μL. Chemically synthesized and modified crRNAs greatly increase the CRISPR-LAMP signal, which advance the wide application. Combined with the previously developed RdRp CRISPR-LAMP assay, clinical results showed that Spike T478K and P681H can discriminate the mutant type form the wild type with 70% (49.66–85.50%, 95% confidence interval) and 78% (57.27–90.62%, 95% confidence interval) sensitivity, respectively, and 100% specificity (51.68–100%, 95% confidence interval), and the RdRp target can detect SARS-CoV-2 strains with 85% sensitivity (65.39–95.14%, 95% confidence interval) and 100% specificity (51.68–100%, 95% confidence interval). We believe that this automatic digital microfluid (DMF) system can advance the integrated CRISPR-LAMP technology with higher stability, sensitivity, and practicability, also for other CRISPR-associated diagnostic platforms.

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