Safe, Effective, and Inexpensive Clearance of Mycoplasma Contamination from Cultures of Apicomplexan Parasites with Sparfloxacin.
Safe, Effective, and Inexpensive Clearance of Mycoplasma Contamination from Cultures of Apicomplexan Parasites with Sparfloxacin.
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DOI：
10.1128/spectrum.03497-22
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发表时间：
2022-10-26
影响因子：
3.7
通讯作者：
中科院分区：
文献类型：
--
作者：
大多数商业产品不能用于清除顶复体寄生虫培养物中的支原体污染，因为寄生虫依赖于顶质体，而顶质体是蓝藻起源的具有DNA复制和翻译机制的重要细胞器。唯一的例外是支原体去除剂（MRA），它相对昂贵，而且一些支原体菌株对MRA的清除表现出耐药性。在这里，我们报道氟喹诺酮类抗生素司帕沙星是一种安全、有效、廉价的治疗顶复体寄生虫培养中支原体污染的替代药物。司帕沙星分别在1 μg/mL和4 μg/mL浓度下清除恶性疟原虫中mra敏感和mra耐药支原体。我们表明，三种不同的顶复合体寄生虫的培养可以维持在清除支原体所需的斯帕沙星浓度下，而不会对寄生虫生长产生实质性的有害影响。我们还描述了另一种低成本的，内部PCR检测支原体。这些发现将有助于实验室在体外维持顶复体寄生虫，特别是在资源匮乏的环境中，在这些环境中，商业产品的高成本为从培养中检测和消除支原体造成了经济障碍。这些发现将有助于实验室在体外维持顶复体寄生虫，特别是在资源匮乏的环境中，在这些环境中，商业产品的高成本为检测和消除培养中的支原体造成了经济障碍。
Most commercial products cannot be used for clearance of Mycoplasma contamination from cultures of apicomplexan parasites due to the parasites’ dependence on the apicoplast, an essential organelle with DNA replication and translation machinery of cyanobacterial origin. The lone exception, mycoplasma removal agent (MRA), is relatively expensive, and some mycoplasma strains have shown resistance to clearance with MRA. Here, we report that the fluoroquinolone antibiotic sparfloxacin is a safe, effective, and inexpensive alternative for treatment of mycoplasma contamination in cultures of apicomplexan parasites. Sparfloxacin cleared both MRA-sensitive and MRA-resistant mycoplasma species from P. falciparum cultures at 1 and 4 μg/mL, respectively. We show that cultures of three different apicomplexan parasites can be maintained at concentrations of sparfloxacin required to clear mycoplasma without resulting in substantial deleterious effects on parasite growth. We also describe an alternative low-cost, in-house PCR assay for detecting mycoplasma. These findings will be useful to laboratories maintaining apicomplexan parasites in vitro, especially in low-resource environments, where the high cost of commercial products creates an economic barrier for detecting and eliminating mycoplasma from culture. IMPORTANCE These findings will be useful to laboratories maintaining apicomplexan parasites in vitro, especially in low-resource environments, where the high cost of commercial products creates an economic barrier for detecting and eliminating Mycoplasma from culture.
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DOI：
10.3791/60985
发表时间：
2020-04-29
期刊：
Journal of visualized experiments : JoVE
影响因子：
--
作者：
Key M;Bergmann A;Micchelli C;Thornton LB;Millard S;Dou Z
通讯作者：
Dou Z
影响因子：
2
作者：
Batiha, Gaber El-Saber;Tayebwa, Dickson Stuart;Igarashi, Ikuo
通讯作者：
Igarashi, Ikuo
影响因子：
4.2
作者：
Gong, Jian-Hua;Liu, Xiu-Jun;Zhen, Yong-Su
通讯作者：
Zhen, Yong-Su
影响因子：
3.7
作者：
Gautier-Bouchardon, Anne V.
通讯作者：
Gautier-Bouchardon, Anne V.
影响因子：
2
作者：
Singh, Shubhra;Puri, S. K.;Srivastava, Kumkum
通讯作者：
Srivastava, Kumkum