Detection of contaminants in cell cultures, sera and trypsin
Detection of contaminants in cell cultures, sera and trypsin
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DOI：
10.1016/j.biologicals.2013.08.005
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发表时间：
2013-11-01
期刊：
影响因子：
1.7
通讯作者：
Heinemann, Marcos Bryan
中科院分区：
文献类型：
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作者：
Pinheiro de Oliveira, Tatiana Flavia;Fonseca, Antonio Augusto, Jr.;Heinemann, Marcos Bryan
本研究旨在标准化和应用聚合酶链反应（PCR）检测细胞培养物、血清和胰蛋白酶中的污染物。标准化了5种PCR方案，以评估细胞培养样品中支原体、猪圆环病毒1 （PCV1）、牛白血病病毒（BLV）或牛病毒性腹泻病毒（BVDV）遗传物质的存在。采用GAPDH和β -actin基因PCR反应评价核酸提取效率。PCR方法应用于来自8个实验室的88个细胞培养样本。这些测试还用于评估来自五个实验室不同批次的10个胰蛋白酶样本和13个胎牛血清样本的潜在污染。结果表明：支原体、PCV1、BVDV RNA和BLV分别为34.1%、35.2%、23.9%和23%。在胎牛血清和胰蛋白酶样品中检测BVDV RNA和PCV1 DNA。结果表明，不同实验室使用的细胞培养物、血清和胰蛋白酶显示出较高的污染物率。研究结果强调了在实验室和使用这些材料的细胞库中监测细胞培养和控制生物污染物的必要性。(C) 2013国际生物标准化联盟。Elsevier Ltd.出版。版权所有。
The aim of this study was standardization and application of polymerase chain reaction (PCR) for the detection of contaminants in cell cultures, sera and trypsin. Five PCR protocols were standardized to assess the presence of genetic material from mycoplasma, porcine circovirus 1 (PCV1), bovine leukemia virus (BLV) or bovine viral diarrhea virus (BVDV) in cell culture samples. PCR reactions for the genes GAPDH and beta-actin were used to evaluate the efficiency of nucleic acid extraction. The PCR protocols were applied to 88 cell culture samples from eight laboratories. The tests were also used to assess potential contamination in 10 trypsin samples and 13 fetal calf serum samples from different lots from five of the laboratories. The results showed the occurrence of the following as DNA cell culture contaminants: 34.1% for mycoplasma, 35.2% for PCV1, 23.9% for BVDV RNA and 23% for BLV. In fetal calf sera and trypsin samples BVDV RNA and PCV1 DNA was detected. The results demonstrated that cell culture, sera and trypsin used by different laboratories show a high rate of contaminants. The results highlight the need for monitoring cell cultures and controlling for biological contaminants in laboratories and cell banks working with these materials. (C) 2013 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.