Objective To establish a sandwich enzyme-linked immunosorbent assay (ELISA) method for the quantitative detection of Chromogranin A (CHGA) in saliva, and to explore its preliminary application in the testing of saliva samples. Methods Recombinant human CHGA protein was used to immunize BALB/c mice, and monoclonal antibodies (mAbs) were prepared and screened using conventional hybridoma technology. A double-antibody sandwich ELISA detection method was constructed, and the matrix effect of saliva samples was optimized. This method was then applied to detect the concentration of CHGA in the saliva of stressed individuals. Results Twenty-one stable hybridoma cell lines secreting high affinity anti-human CHGA antibodies were obtained. A pair of detection antibodies with the best effect was selected, and the optimal coating concentration was determined to be 10 μg/mL, with the optimal dilution of detection antibodies being 1:32 000. The accuracy and reproducibility of this method were verified, with both intra-batch and inter-batch variation coefficients less than 15×, and the recovery rate between 80× and 120×. The matrix effect was further optimized to make it suitable for saliva sample detection. Saliva samples from individuals in different stress states were collected, and the CHGA levels were detected using the method established in this study, indicating its potential to reflect the intensity of stress. Conclusion A reliable saliva CHGA ELISA detection method has been successfully established, and its potential as a biomarker in stress-related research has been preliminarily explored.
Saliva/metabolism* ; Enzyme-Linked Immunosorbent Assay/methods* ; Humans ; Animals ; Mice, Inbred BALB C ; Mice ; Chromogranin A/immunology* ; Antibodies, Monoclonal/immunology* ; Female ; Male ; Reproducibility of Results ; Adult

At present, there are many problems in the nail-fold microcirculation detection devices, such as huge structure, inconvenience to carry. In addition, due to the patient's body shaking, the video is difficult to keep stable in collecting with the device, which brings great difficulties to the doctor's observation. We develop a small image acquisition device for nail-fold microcirculation based on the principle of SDF imaging principle and liquid lenses technology. An annular lighting device is fixed in front of the optical system, and the overall design of the system is based on the characteristics of human fingers. The device is small, easy to carry and conform to the fingertips. It can focus quickly through a controller. It can also achieve high quality images of the nail-fold microcirculation. This study can promote the usage of nail-fold microcirculation device at the bedside. It's an efficient tool for medical workers to observe the microcirculation of patients.
Humans ; Microcirculation