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目的研制可提高荒漠型黑热病婴儿利什曼原虫分离率的培养基。方法荒漠型黑热病患者的感染材料,分别接种3N培养基、RPMI-1640完全培养基、配方1、配方2和配方3培养基,观察比较利什曼原虫在5种培养基的培养、分离、传代和保存效果。比较5个温度条件下前鞭毛体在配方3的生长曲线。结果婴儿利什曼原虫在5种培养基培养13d的培养效果基本一致,第3d出现前鞭毛体,第7d达到高峰,第13d几乎全部凋亡。用5种培养基每7d传代培养1次,除配方3能延续传代保存前鞭毛体外,其他4种培养基在传代培养过程中前鞭毛体逐渐丢失。用配方3培养前鞭毛体,20~30℃在第6d出现生长峰值;20℃在第20d出现第二个峰值;10℃和14℃在12d和第20d分别出现第一个和第二个峰值,最长可维持到第37d。结论 3N培养基、RPMI-1640完全培养基、配方1、配方2和配方3培养基均适用于婴儿利什曼原虫的前期培养,第3d就可以判断是否有前鞭毛体生长,但只有配方3培养基具有延续传代培养婴儿利什曼原虫的效果。
Objective To develop a medium that can improve the isolation rate of Leishmania babes in desert kala-azar. Methods Infected materials of patients with desert kala azar were inoculated with 3N medium, RPMI-1640 complete medium, Formulation 1, Formulation 2 and Formulation 3 medium respectively. The culture, isolation and passage of Leishmania in 5 kinds of medium were observed and compared And save the effect. The growth curve of Promastigotes in Formulation 3 was compared under five temperature conditions. Results The culture effect of Leishmania infantum on the 13th day of culture was basically the same. The promastigotes appeared on the 3dth day and peaked on the 7th day. Almost all of them died on the 13th day. With 5 kinds of medium every 7d subculture 1 time, in addition to the formula 3 can continue pre-passage preservation pre-flagella body, the other 4 kinds of medium during subculturing promastigotes gradually lost. The pre-cultured flagella were cultured in the formula 3, the peak appeared at 20 ~ 30 ℃ on the 6th day, the second peak appeared on the 20th day at 20 ℃, and the first and second peak appeared on the 12th and the 20th respectively at 10 ℃ and 14 ℃ , Up to the first 37d. Conclusion 3N medium, RPMI-1640 complete medium, Formulation 1, Formulation 2 and Formulation 3 medium are all suitable for the pre-culture of Leishmania infantum. The first 3d can judge whether there is promastigote growth, but only Formulation 3 The medium has the effect of prolonging subculturing Leishmania infantum.