基于环介导等温扩增技术的空间微生物安全水平评价方法

Evaluation method for safety level of space microbial based on the loop-mediated isothermal amplification

以金黄色葡萄球菌、肺炎克雷伯菌、青霉、黑曲霉4种典型空间微生物风险菌为试验对象,在筛选得到靶标微生物的特异性环介导等温扩增(LAMP)引物的基础上,成功建立基于LAMP的靶标微生物定量检测技术,检出限为200 copies/μL。该方法表现出良好的检测灵敏度、检测特异性和检测精密度,为空间微生物安全水平评价提供了一种切实可行的方法,为保障航天员健康和人机系统的安全性提供了重要的技术手段。

MoS_(2) doped carbon nanoenzymes(Mo,S-CDs) was synthesized by hydrothermal method with sodium lignosulfonate as carbon source, and applied to detect the content of dopamine in human urine based on its peroxidase characteristics. The morphology of nanoenzyme was characterized. The results showed that Mo,S-CDs were spherical in shape with a diameter of about 2 nm and could be well dispersed in water. The infrared spectrum proved that there were abundant functional groups on the surface of Mo,S-CDs, and X-ray photoelectron spectroscopy(XPS) proved that Mo, S, C and O existed in Mo,S-CDs. Mo,S-CDs have stable and efficient peroxidase catalytic activity, which can catalyze the reaction between H_(2)O_(2) and 3,3',5,5'-tetramethylbenzidine(TMB) to produce oxTMB. The blue color oxTMB can be reduced to TMB by the reductive dopamine, resulting in the decrease of the system absorbance at the characteristic peak of oxTMB. Therefore, the catalytic sensing system of dopamine Mo,S-CDs were constructed based on the sensing ability of Mo,S-CDs to H_(2)O_(2), which could detect the content of dopamine in human urine. There was a good linear relationship in the dopamine concentration range of 0.5-20 μmol/L, and the detection limit was 0.0639 μmol/L. The recoveries were 96.5%-101.7%, and the relative standard deviations(RSDs) were less than 5%.