A multi-laboratory profile of Mycoplasma contamination in Lawsonia intracellularis cultures.
A multi-laboratory profile of Mycoplasma contamination in Lawsonia intracellularis cultures.
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DOI：
10.1186/1756-0500-5-78
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发表时间：
2012-01-27
影响因子：
1.8
通讯作者：
Yeh JY
中科院分区：
文献类型：
--
作者：
Hwang JM;Lee JH;Yeh JY
在实验室常规培养过程中，支原体污染一直是一个常见的问题。当研究胞内乳杆菌的实验室出现支原体污染时，必须丢弃培养物，原因有四：1)在胞内乳杆菌传代过程中，支原体不可避免地与胞内乳杆菌一起聚集；2)支原体抑制胞内乳杆菌的生长；3)胞内乳杆菌培养物中的支原体不可能被选择性地清除。在这项研究中，我们观察了多个实验室培养胞内乳杆菌过程中支原体的污染情况。用聚合酶链式反应(PCR)检测培养物中是否存在支原体感染，并对部分16S rRNA和23S rRNA基因进行了序列分析。用属特异性通用引物进行的聚合酶链式反应检测发现，34株培养物中有29株(85.3%)被支原体污染，其中猪鼻支原体26株(89.2%)，口腔支原体2株(6.9%)，发酵支原体1株(3.4%)。支原体污染不是由猪源材料感染造成的。培养胞内乳杆菌所需的麦考伊细胞也被排除为支原体污染源。在这项研究中，猪鼻分枝杆菌被鉴定为最常见的污染胞内乳杆菌培养物。胞内乳杆菌是否能增强支原体的生物学特性以促进McCoy细胞的感染尚不清楚。由于同时使用的McCoy细胞系的菌种均为支原体阴性，而且同一工人同时处理McCoy细胞以保持细菌和胞内乳杆菌的培养，因此胞内乳杆菌培养物可能更容易受到支原体的污染。综上所述，这些结果表明，连续培养的胞内乳杆菌必须定期检测支原体污染。本文报道的序列的GenBank登录号为JN689375至JN689377。
During the routine laboratory cultivation of Lawsonia intracellularis, Mycoplasma contamination has been a frequent problem. When Mycoplasma contamination occurs in laboratories that study L. intracellularis, the cultures must be discarded for 4 reasons: 1) Mycoplasma is inevitably concentrated along with L. intracellularis during the passage of L. intracellularis; 2) Mycoplasma inhibits the growth of L. intracellularis; and 3) it is impossible to selectively eliminate Mycoplasma in L. intracellularis cultures. In this study, we observed the contamination of Mycoplasma species during L. intracellularis cultivation among multiple laboratories. The presence of a Mycoplasma infection in the L. intracellularis cultures was verified using polymerase chain reaction (PCR), and a sequence analysis of the partial 16S rRNA and 23S rRNA genes was performed. A PCR-based assay using genus-specific universal primers revealed that 29 (85.3%) of the 34 cultures were contaminated with Mycoplasma, including 26 with M. hyorhinis (89.2%), 2 with M. orale (6.9%), and 1 with M. fermentans (3.4%). The Mycoplasma contamination was not the result of infection with material of pig origin. McCoy cells, which are required for the cultivation of L. intracellularis, were also ruled out as the source of the Mycoplasma contamination. In this study, M. hyorhinis was identified as the most common mollicute that contaminated L. intracellularis cultures. Whether L. intracellularis enhances the biological properties of Mycoplasma to promote infection in McCoy cells is not known. Because the McCoy cell line stocks that were used simultaneously were all negative for Mycoplasma, and the same worker handled both the McCoy cells to maintain the bacteria and the L. intracellularis cultures, it is possible that the L. intracellularis cultures are more vulnerable to Mycoplasma contamination. Taken together, these results suggest that continuous cultures of L. intracellularis must be tested for Mycoplasma contamination at regular intervals. The GenBank accession numbers for the sequences reported in this paper are JN689375 to JN689377.