Theunissen J J, Stolz E, Michel M F
Department of Dermato-Venereology, Erasmus University, Rotterdam, The Netherlands.
J Appl Bacteriol. 1993 Nov;75(5):473-7. doi: 10.1111/j.1365-2672.1993.tb02804.x.
The effects of suspension media and rate of freezing on the survival of Chlamydia trachomatis LGV2 and Chlamydia pneumoniae after lyophilization were assessed. The highest loss in infectious elementary bodies (EBs) occurred during lyophilization. The survival was higher after freezing at a rate of 1 degree C min-1 and lyophilization than that after rapid freezing at -70 degrees C or -196 degrees C. The recovery (+/- 5%) was higher when fetal calf serum (FCS) containing glucose, saccharose or lactose were used as lyophilization media than that (0.5-3%) when yolk-sac, skimmed milk or phosphate buffer containing sucrose, glutamine and 10% FCS (SPG) were used. After lyophilization, the survival was not affected in the tested range from 10(4) to 5 x 10(6) inclusion-forming units (ifu) ml-1 prior to freezing. After storage for 4 months at 4 degrees C, the numbers of ifu of both Chlamydia serovars that were recovered were identical to the numbers of ifu immediately after lyophilization. It was concluded that chlamydias can be stored and transported in lyophilized form. However, a loss of 95% in infectious EBs should be taken into account.
评估了悬浮介质和冷冻速率对冻干后沙眼衣原体LGV2和肺炎衣原体存活的影响。冻干过程中感染性原体(EBs)损失最大。以1℃/分钟的速率冷冻后再冻干,其存活率高于在-70℃或-196℃快速冷冻后的存活率。当使用含有葡萄糖、蔗糖或乳糖的胎牛血清(FCS)作为冻干介质时,回收率(±5%)高于使用含有蔗糖、谷氨酰胺和10%FCS的卵黄囊、脱脂牛奶或磷酸盐缓冲液(SPG)时的回收率(0.5 - 3%)。冻干后,冷冻前每毫升10⁴至5×10⁶个包涵体形成单位(ifu)的测试范围内,存活率不受影响。在4℃储存4个月后,两种衣原体血清型回收的ifu数量与冻干后立即回收的ifu数量相同。得出的结论是衣原体可以以冻干形式储存和运输。然而,应考虑到感染性EBs损失95%的情况。