MycAwayTMPlus-Color One-Step Mycoplasma Detection Kit是利用Yeasen独特的等温扩增技术开发的针对细胞培养液中支原体污染的快速检测产品，较以往支原体检测产品本试剂盒做了升级改善，在极大程度上降低了假阳性率，提高检测的准确度并增强了阴性和阳性的辨识度。在扩增结束后，可在室温放置一段时间且不会出现由阴性转变为阳性的现象，不影响阴、阳性判断。主要原理是若细胞培养物被支原体污染，支原体DNA的保守序列会被大量、快速地扩增，使反应液由蓝紫色变成天蓝色，结果肉眼可辨，无需电泳。
太阳成集团MycAwayTMPlus-Color One-Step Mycoplasma Detection Kit可以检测多种支原体，包括细胞培养过程中常见的8种支原体。传统巢式PCR支原体法检测法易受细胞培养上清中抑制物的影响，产生假阴性结果；反应后需开盖电泳检测，增加污染导致的假阳性风险。一步法支原体检测试剂盒完全没有上述缺点，且其检测灵敏度、准确性远远高于PCR法。
1、检测种类多：可精确检测22种支原体，包括常见8种支原体；
2、结果分辨性强：检测结果参照标准清晰，检出率高，阴、阳性分辨性强；；
3、检测性能强：通过与竞品对比，试剂盒在63℃情况下从30min、40min到50min，稳定性远高于竞品，且无假阳性；
4、反应时长极限长：反应放置时长可以从60min、90min到120min均无弱阳性出现，且NTC与原始颜色无明显差别；
5、反应结束室温放置稳定性强： 63℃反应结束后，无需95℃灭活，可直接放置于室温3天，阴阳性结果与原始无明显差别。
检测性能比较
结论：
1）竞品一30min检出为10万；在40min出现NTC转阳现象；50min完全转阳；无法分辨阴性与阳性；假阳性率高。
2）竞品二30min检出为1000copies/μL，但伴随NTC出现弱阳性，且出现显色混乱，无法分辨阴性与阳性；40min完全混乱，假阳性率极高。
3）相较于竞品，40612ES 30min检出均与竞品一或竞品二相当，但其稳定性远高于竞品，无假阳性出现。
本次测试采用支原体DNA为模板，投入为1uL。
冰袋运输。-25 ~ -15℃避光保存，保质期18个月。若较长时间不用，请注意避光保存。
Q：40612 一步法快速支原体检测试剂盒（防污染版）的检测原理？
A：本试剂盒中加入了UDG 酶，以便去除含 dU 的PCR 产物气溶胶污染，在极大程度上抑制假阳性反应，提高检测的准确度。而且加入的 UDG 酶在室温下起作用，且对温度感、易于灭活，不影响实验结果。若细胞培养物被支原体污染，支原体 DNA 的保守序列会被大量、快速地扩增，使反应液由蓝紫色变成天蓝色，结果肉眼可辨，无需电泳。
Q：一步法快速支原体检测试剂盒使用方法（防污染版）？
A：本试剂盒使用更为简便，只需吸取 1μL 疑似支原体污染的细胞培养上清加入反应体系中，先将样品在 25~37 ℃放置 5 min，再在水浴锅或 PCR 仪中，恒温 63 °C 孵育 60 min。观察反应液由蓝紫色变成天蓝色，结果肉眼可辨，无需电泳。
Q：用 40612 一步法支原体检测试剂盒检测 DMSO、乙醇等有机溶剂中是否存在支原体污染，发现实验组均是天蓝色，是不是 DMSO、乙醇等时机中存在支原体污染？
A：支原体试剂盒在乙醇、DMSO 有机溶剂，以及强酸强碱环境中，不稳定易变性，导致颜色变色为天蓝色。这款试剂盒只能用于细胞检测，不可用于有机溶剂、强酸强碱环境下检测。
Q：我检测的是这个里面的液体，加入之后PCR管里的颜色立马变成了天蓝色？是什么原因呢？检测胰酶-EDTA，加入样品之后，直接变色了？
A：40612原理：Mg2++HNB=蓝紫色，随着PCR增加，焦磷酸不断增多，焦磷酸与Mg2+结合，HNB游离，显示天蓝色。这个试剂里面有磷酸二氢钠可能是这个离子与Mg2+结合而立马产生化学反应导致的，EDTA可以螯合钙镁离子，从而导致反应加速发生变色反应。
Q：阳性对照的浓度时多少？
A：10的5次方copies/ul。
Q：阳性对照里的是支原体吗？
A：不是的，是线性化的片段。
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