Pickles S F, Pritchard D I
Division of Regenerative Medicine and Cellular Therapies, School of Pharmacy, University of Nottingham, Nottingham, U.K.
Med Vet Entomol. 2017 Jun;31(2):200-206. doi: 10.1111/mve.12220. Epub 2017 Jan 24.
Lucilia sericata Meigen (Diptera: Calliphoridae) larvae are manufactured worldwide for the treatment of chronic wounds. Published research has confirmed that the primary clinical effect of the product, debridement (the degradation of non-viable wound tissue), is accomplished by a range of enzymes released by larvae during feeding. The quality assessment of larval activity is currently achieved during production using meat-based assays, which monitor insect growth and/or the reduction in substrate mass. To support this, the present authors developed a complementary radial diffusion enzymatic assay to produce a visual and measureable indication of the activity of larval alimentary products (LAP) collected under standardized conditions, against a gelatin substrate. Using basic laboratory equipment and reagents, the assay is rapid and suited to high throughput. Assay reproducibility is high (standard deviation: 0.06-0.27; coefficient of variation: 0.75-4.31%) and the LAP collection procedure does not adversely affect larval survival (mortality: < 2%). Because it is both cost- and time-effective, this method is suited to both academic and industrial use and supports good manufacturing and laboratory practice as a quality control assay.
丝光绿蝇（双翅目：丽蝇科）幼虫在全球范围内被用于生产治疗慢性伤口的产品。已发表的研究证实，该产品的主要临床效果——清创（即坏死伤口组织的降解），是由幼虫进食期间释放的一系列酶来实现的。目前，幼虫活性的质量评估是在生产过程中使用基于肉类的检测方法进行的，这些方法监测昆虫的生长和/或底物质量的减少。为了辅助这一评估，本文作者开发了一种补充性的径向扩散酶检测方法，以针对明胶底物，对在标准化条件下收集的幼虫消化产物（LAP）的活性给出直观且可测量的指示。该检测方法使用基本的实验室设备和试剂，操作快速且适合高通量检测。检测的重现性很高（标准差：0.06 - 0.27；变异系数：0.75 - 4.31%），并且LAP收集过程不会对幼虫存活产生不利影响（死亡率：<2%）。由于该方法具有成本效益且省时，因此适用于学术和工业用途，并作为一种质量控制检测方法支持良好的生产和实验室规范。
