Wood D E, Smrkovski L L, McConnell E, Pacheco N D, Bawden M P
Bull World Health Organ. 1979;57 Suppl 1(Suppl):69-74.
An improved procedure is presented for the isolation of Plasmodium berghei sporozoites from host mosquitos. The method employs filtration through a series of Nuclepore membranes followed by two consecutive centrifugations of the filtrate layered over Renografin-60 solutions of different densities. A Coulter Counter was used to compare isolations prepared by this technique with those prepared by a routinely employed discontinuous gradient method. When the sporozoite concentration in each preparation was standardized at 300 sporozoites per ml, isolations prepared by the new technique were significantly cleaner than isolations prepared by the discontinuous gradient method, containing an average of 1706 total particles per ml compared with 46 107 total particles per ml. The latter procedure was more effective, however, in removing viable microorganisms. Sporozoites isolated by both techniques were similar in immunogenicity and virulence.
本文介绍了一种从宿主蚊子中分离伯氏疟原虫子孢子的改进方法。该方法采用通过一系列核孔膜过滤，然后将滤液在不同密度的泛影葡胺 - 60溶液上分层进行两次连续离心。使用库尔特计数器将用该技术制备的分离物与通过常规使用的不连续梯度法制备的分离物进行比较。当每种制剂中的子孢子浓度标准化为每毫升300个子孢子时，新技术制备的分离物比不连续梯度法制备的分离物明显更纯净，每毫升平均含有1706个总颗粒，而后者为每毫升46107个总颗粒。然而，后一种方法在去除活微生物方面更有效。通过两种技术分离的子孢子在免疫原性和毒力方面相似。
