液体培养基分离和保存利什曼原虫的效果观察

Effects of liquid medium on the isolation and preservation of Leishmania spp

目的观察利什曼原虫液体培养基(LLM)分离和保存利什曼原虫的效果。方法用LLM培养基对黑热病材料进行利什曼原虫分离培养,同时接种的灰仓鼠,进行利什曼原虫分离率比较。结果用LLM培养被检材料出结果时间为4d,对荒漠型和平原型黑热病利什曼原虫分离率分别为35.7%(10/28)和50.0%(15/30),分别比接种灰仓鼠的分离率高10%和30%。培养基-20℃冷冻保存有效期为2年。结论应用LLM培养基能显著提高利什曼原虫的分离率。该培养基与传统3N培养基比较,保存时间延长,培养出结果时间缩短,可用于黑热病实验诊断和黑热病抗药性研究。

Objective To observe the effects of liquid medium（LM）on the isolation and culturing of Leishmania spp.Methods LM was used to isolate and culture Leishmania spp.Grey hamsters（Cricetulus migratorius）were also infected with Leishmania spp.and the rate of Leishmania isolation was compared. Results Leishmania specimens were cultured on LM for 4days.Leishmania responsible for zoonotic visceral leishmaniasis was isolated at a rate of 35.7%（10/28）and Leishmania responsible for anthroponotic visceral leishmaniasis was isolated at a rate of 50%（15/30）.In contrast,infection of grey hamsters resulted in isolation of Leishmania responsible for zoonotic visceral leishmaniasis at a rate of 10%and isolation of Leishmania responsible for anthroponotic visceral leishmaniasis at a rate of 30%.Leishmania spp.cultured on LM were effectively cryopreserved at-20℃for two years. Conclusion LM significantly improved the isolation of Leishmania spp.In comparison to conventional 3N medium,LM allowed a longer duration of preservation（two years instead of one month）and it yielded results sooner（15days instead of four days）.LM can be used for the laboratory diagnosis of visceral leishmaniasis and for study of its drug resistance.