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冷冻保存可降低受污染的外周血祖细胞产品中可检测到的细菌浓度。

Cryopreservation reduces the concentration of detectable bacteria in contaminated peripheral blood progenitor cell products.

作者信息

Kipp Frank, Linnemann Elmar, Fischer Rudolf-Josef, Sibrowski Walter, Cassens Uwe

机构信息

Institute of Medical Microbiology, the Institute of Transfusion Medicine/Transplantation Immunology, and the Department of Biometrics, University Hospital Münster, Germany.

出版信息

Transfusion. 2004 Jul;44(7):1098-103. doi: 10.1111/j.1537-2995.2004.03392.x.

Abstract

BACKGROUND

Microbial contamination of PBPC products PBPCPs may cause severe clinical complications. There-fore, we investigated the influence of cryopreservation on the sensitivity to detect bacterial contaminations in PBPCPs.

STUDY DESIGN AND METHODS

Expired PBPCPs (n = 29) were thawed, and defined concentrations of Staphylococcus epidermidis or Escherichia coli were inoculated into each bag. After 60 minutes of intermixing, a representative aliquot was drawn and cultured on Mueller-Hinton agar for 24 hours. Then, the products were cryopreserved for 24 hours, and the procedure was repeated as mentioned above. The total numbers of CFUs were counted before and after cryopreservation.

RESULTS

A mean concentration of 2529 CFUs per mL of S. epidermidis was determined before cryopreservation versus 2182 CFUs per mL after cryopreservation, demonstrating a decrease of detectable colonies (p < 0.05). For E. coli, the mean numbers were 424 CFUs per mL before cryopreservation and 343 CFUs per mL after cryopreservation, also showing a decrease (p < 0.05).

CONCLUSION

The cryopreservation reduces the concentration of detectable bacteria in contaminated PBPCPs. Especially in sterility testing of PBPCPs with low bacterial contamination, this phenomenon could lead to false-negative results with severe clinical consequences.

摘要

背景

外周血祖细胞产品(PBPCPs)的微生物污染可能会导致严重的临床并发症。因此,我们研究了冷冻保存对外周血祖细胞产品中细菌污染检测灵敏度的影响。

研究设计与方法

将过期的外周血祖细胞产品(n = 29)解冻,向每个袋子中接种规定浓度的表皮葡萄球菌或大肠杆菌。混合60分钟后,抽取一份代表性的等分试样,在穆勒-欣顿琼脂上培养24小时。然后,将产品冷冻保存24小时,并按上述方法重复该过程。在冷冻保存前后计算菌落形成单位(CFU)的总数。

结果

冷冻保存前表皮葡萄球菌的平均浓度为每毫升2529个CFU,冷冻保存后为每毫升2182个CFU,表明可检测菌落数量减少(p < 0.05)。对于大肠杆菌,冷冻保存前的平均数量为每毫升424个CFU,冷冻保存后的平均数量为每毫升343个CFU,也呈现出减少趋势(p < 0.05)。

结论

冷冻保存会降低受污染外周血祖细胞产品中可检测到的细菌浓度。尤其是在外周血祖细胞产品细菌污染程度较低的无菌检测中,这种现象可能会导致假阴性结果,从而产生严重的临床后果。

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