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杜氏利什曼原虫前鞭毛体在血琼脂斜面中的保存

Preservation of Leishmania donovani promastigotes in blood agar slants.

作者信息

Muniaraj M, Gupta A K, Narayan S, Singh D, Sinha P K, Kishore K, Das P, Bhattacharya S K

机构信息

Rajendra Memorial Research Institute of Medical Sciences, (ICMR) Agam Kuan, Patna.

出版信息

J Commun Dis. 2005 Sep;37(3):191-5.

Abstract

Long-term cultivation of Leishmania promastigotes by weekly passage to fresh medium was reported to be disadvantageous because needs labor, risk of contamination, lowering in infectivity and virulence pattern. Cryopreservation and Lyophilization require expensive facilities which could be a burden and unaffordable to most laboratories of developing countries where the disease is endemic. These problems could be minimized by simple preservation of Leishmania donovani promastigotes in blood agar slants at 7-8 degrees C for 6-7 months. The preserved promastigotes were examined for viability up to one year at a regular interval of one month. Viable promastigotes were found and revived successfully from all the slants stored up to 7 months after that, the viability of promastigotes was found to be decreased in the slants of 8-9 month storage. No viable promastigotes were recovered from the slants stored up to 11-12 months. By this method, the promastigotes can easily be stored up to 7 months without loss of biological activity. The number of passage of promastigotes to fresh medium has been greatly reduced by this method from 30 times to 01 when compared with weekly passage in liquid medium. This simple and economical method can be recommended for short storage of Leishmania culture without loss of any activity.

摘要

据报道,通过每周转接至新鲜培养基对利什曼原虫前鞭毛体进行长期培养存在弊端,因为这需要人工操作、有污染风险、感染力和毒力模式会降低。冷冻保存和冻干需要昂贵的设备,这对于疾病流行的大多数发展中国家的实验室来说可能是一种负担且难以承受。通过将杜氏利什曼原虫前鞭毛体简单保存在7 - 8摄氏度的血琼脂斜面上6 - 7个月,这些问题可以得到最小化。每隔一个月定期检查保存的前鞭毛体长达一年的活力。在保存长达7个月的所有斜面中都发现了有活力的前鞭毛体并成功复苏,之后发现在保存8 - 9个月的斜面中前鞭毛体的活力下降。在保存长达11 - 12个月的斜面中未回收到有活力的前鞭毛体。通过这种方法,前鞭毛体可以轻松保存长达7个月而不丧失生物活性。与在液体培养基中每周转接相比,通过这种方法,前鞭毛体转接至新鲜培养基的次数已从30次大幅减少至1次。这种简单且经济的方法可推荐用于利什曼原虫培养物的短期保存而不丧失任何活性。

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