Combined detection of SARS-CoV-2 neutralizing antibodies and specific IgG in plasma based on SERS magnetic sensor.

In this study, a surface-enhanced Raman spectroscopy (SERS) magnetic sensor is established based on SERS principle and magnetic separation technology, and a highly sensitive, simple and fast method for quantitative detection of neutralizing antibodies (nABs) and specific IgG of SARS-CoV-2 in plasma is established combined with immunoassay. Two kinds of Raman nanospheres (RNPs) with different characteristic Raman shifts are used as signal sources and coupled to ACE2 and anti-IgG (FC) antibodies respectively, and magnetic beads are coupled to RBD. The competitive relationship between ACE2 and nABs, the binding relationship between specific IgG and anti-IgG (FC) antibodies are determined. The results show that the concentrations of nABs and specific IgG in the range of 10-2000 ng ml-1are well correlated with SERS response intensity, and the recoveries are both between 90%-110%, with good precision. Bilirubin and common anticoagulants have no interference on the detection results. This method is accurate, reliable, sensitive and does not require complex pre-treatment, and is expected to be used for simultaneous detection of nABs and specific IgG in plasma of SARS-CoV-2. It has guiding significance for the development and evaluation of vaccines and the formulation of individualized vaccination schedule.

Development of a system for detecting cardiac troponin I by background fluorescence quenching based on internal filtration effect.

The present study sought to develop a cardiac troponin I (cTnI) detection system based on background fluorescence quenching of internal filtration effect (IFE) and study the influence of IFE on the sensitivity of cTnI detection. Three nanogold materials were synthesized as fluorescence quenchers, and rhodamine 6 G (R6G) and Cy5 were used as fluorescence probes. Six experimental systems were established to detect cTnI in negative serum test solutions and clinical serum samples. The sensitivity of each system was compared to explore the contribution of IFE to the detection sensitivity of cTnI. When applied to negative serum test solutions, the R6G-nanogold material I system exhibited a superior detection effect for cTnI, with a limit of detection (LOD) of 0.15 ng ml-1. When applied to clinical serum samples, the Cy5-nanogold material Ⅲ system yielded a better detection effect for cTnI, with the lowest concentration of cTnI detected at 2 ng ml-1. The first and second internal filtering effects in the proposed system can be achieved simultaneously, effectively avoiding light absorption interference from clinical serum samples and enhancing the sensitivity of the background fluorescence quenching detection of cTnI.

Study on the performance of novel nanomaterials for detection of biomarkers such as PCT based on immunochromatography sensitivity.

In this study, by comparing the UV-vis spectral characteristics of colloidal gold and colloidal gold enhancer, and their differences as immunochromatographic tracers in the qualitative detection of PCT, IL-6, Hp and quantitative determination of PCT performance, the factors that may affect the sensitivity were discussed. The results show that the absorbance at 520 nm of CGE diluted 20-fold and colloidal gold diluted 2-fold were comparable, and the sensitivity of CGE immunoprobe for qualitative detection of PCT, IL-6 and Hp was higher than that of colloidal gold immunoprobe, and the reproducibility and accuracy of both immunoprobes for quantitative detection of PCT were good. Indicating that the high sensitivity of CGE immunoprobe detection is mainly due to the absorption coefficient of CGE at 520 nm is about 10 times that of colloidal gold immunoprobe, CGE has stronger light absorption capacity and stronger quenching effect on rhodamine 6 G on the nitrocellulose membrane surface of the test strip.

Synchronous detection of IgG subtypes based on SERS combined with immunoassay.

In this study, a rapid, simple, highly sensitive and anti-interference method for the joint detection of four IgG subtypes is established by using Raman microspheres with four characteristic Raman spectra. The results show that the concentrations of IgG1 in the range of 0-1500 ng ml-1, IgG2 in the range of 0-1100 ng ml-1, IgG3 in the range of 0-88.7 ng ml-1, IgG4 in the range of 0-77.2 ng ml-1, it shows a good correlation with the response value of The Raman signal. The lowest detection limits are 25.4 ng ml-1, 21.7 ng ml-1, 1.6 ng ml-1, 1.7 ng ml-1, respectively. Reproducibility is good, the coefficient of variation of low, medium and high concentration standard solution are within 10%. The recoveries of four IgG subtypes are in the range of 90%-110%, and the accuracy of the method is good. The coefficients of variation between and within the three batches of reagents are all less than 11%, showing good precision. There is no cross reaction with Procalcitonin (20 ng ml-1), Interleukin-6 (1 ng ml-1) and bovine serum albumin (10 mg ml-1), and the specificity is good. Common interfering substances such as bilirubin, triglyceride and trisodium citrate do not affect the determination results, and heparin sodium only affects the determination results of IgG1. This method has good anti-interference ability. The method has high sensitivity, simple operation and strong anti-interference ability, and has good correlation with the IgG detection methods commonly used in clinic. This simple and quantitative method can be used for the rapid detection of IgG subtypes in the future, which can improve the efficiency of clinical diagnosis.

Role of glutamine in the mediation of E-cadherin, p120-catenin and inflammation in ventilator-induced lung injury.

BACKGROUND: Ventilator-induced lung injury (VILI) is commonly associated with barrier dysfunction and inflammation reaction. Glutamine could ameliorate VILI, but its role has not been fully elucidated. This study examined the relationship between inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor [TNF]-α, and IL-10) and adherens junctions (E-cadherin, p120-catenin), which were ameliorated by glutamine in VILI, both in vitro and in vivo. METHODS: For the in vivo study, 30 healthy C57BL/6 mice weighing 25-30 g were randomly divided into five groups with random number table (n = 6 in each group): control (Group C); low tidal volume (Group L); low tidal volume + glutamine (Group L + G); high tidal volume (Group H); and high tidal volume + glutamine (Group H + G). Mice in all groups, except Group C, underwent mechanical ventilation for 4 h. For the in vitro study, mouse lung epithelial 12 (MLE-12) cells pretreated with glutamine underwent cyclic stretching at 20% for 4 h. Cell lysate and lung tissue were obtained to detect the junction proteins, inflammatory cytokines, and lung pathological changes by the Western blotting, cytokine assay, hematoxylin and eosin staining, and immunofluorescence. RESULTS: In vivo, compared with Group C, total cell counts (t = -28.182, P < 0.01), the percentage of neutrophils (t = -28.095, P < 0.01), IL-6 (t = -28.296, P < 0.01), and TNF-α (t = -19.812, P < 0.01) in bronchoalveolar lavage (BAL) fluid, lung injury scores (t = -6.708, P < 0.01), and the wet-to-dry ratio (t = -15.595, P < 0.01) were increased in Group H; IL-10 in BAL fluid (t = 9.093, P < 0.01) and the expression of E-cadherin (t = 10.044, P < 0.01) and p120-catenin (t = 13.218, P < 0.01) were decreased in Group H. Compared with Group H, total cell counts (t = 14.844, P < 0.01), the percentage of neutrophils (t = 18.077, P < 0.01), IL-6 (t = 18.007, P < 0.01), and TNF-α (t = 10.171, P < 0.01) in BAL fluid were decreased in Group H + G; IL-10 in BAL fluid (t = -7.531, P < 0.01) and the expression of E-cadherin (t = -14.814, P < 0.01) and p120-catenin (t = -9.114, P < 0.01) were increased in Group H + G. In vitro, compared with the nonstretching group, the levels of IL-6 (t = -21.111, P < 0.01) and TNF-α (t = -15.270, P < 0.01) were increased in the 20% cyclic stretching group; the levels of IL-10 (t = 5.450, P < 0.01) and the expression of E-cadherin (t = 17.736, P < 0.01) and p120-catenin (t = 16.136, P < 0.01) were decreased in the 20% cyclic stretching group. Compared with the stretching group, the levels of IL-6 (t = 11.818, P < 0.01) and TNF-α (t = 8.631, P < 0.01) decreased in the glutamine group; the levels of IL-10 (t = -3.203, P < 0.05) and the expression of E-cadherin (t = -13.567, P < 0.01) and p120-catenin (t = -10.013, P < 0.01) were increased in the glutamine group. CONCLUSIONS: High tidal volume mechanical ventilation and 20% cyclic stretching could cause VILI. Glutamine regulates VILI by improving cytokines and increasing the adherens junctions, protein E-cadherin and p120-catenin, to enhance the epithelial barrier function.