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储存温度和时间对采用空斑小室培养法和聚合酶链反应技术检测血液标本中巨细胞病毒的定性和定量结果的影响。

Effects of storage temperature and time on qualitative and quantitative detection of cytomegalovirus in blood specimens by shell vial culture and PCR.

作者信息

Roberts T C, Buller R S, Gaudreault-Keener M, Sternhell K E, Garlock K, Singer G G, Brennan D C, Storch G A

机构信息

Division of Infectious Diseases, Washington University School of Medicine, St. Louis, Missouri 63110, USA.

出版信息

J Clin Microbiol. 1997 Sep;35(9):2224-8. doi: 10.1128/jcm.35.9.2224-2228.1997.

Abstract

Cytomegalovirus (CMV) infectious titers and DNA levels were determined by quantitative shell vial culture and quantitative-competitive PCR with blood samples from 10 renal transplant recipients with active CMV infection. Blood samples were stored at either room temperature or 4 degrees C and were processed at intervals of 0, 6, 24, 48, and 72 h. All samples were culture and PCR positive at baseline. Whereas the sensitivity of shell vial culture progressively declined, with only 55% positive at 24 h and 10% positive at 48 h, all samples remained PCR positive at all time points. Furthermore, the infectious titer diminished by 83 to 91% by 24 h compared to that at baseline (P < 0.0001), but quantitative DNA levels did not decline over time. Storage temperature had no significant effect on either infectious titer or DNA levels.

摘要

采用定量空斑培养法和定量竞争聚合酶链反应(PCR),对10例有活动性巨细胞病毒(CMV)感染的肾移植受者的血样进行检测,以确定CMV感染滴度和DNA水平。血样分别储存在室温或4℃条件下,并在0、6、24、48和72小时的间隔时间进行处理。所有样本在基线时培养和PCR检测均为阳性。空斑培养的敏感性逐渐下降,24小时时只有55%为阳性,48小时时为10%阳性,而所有样本在所有时间点PCR检测均保持阳性。此外,与基线时相比,24小时时感染滴度降低了83%至91%(P<0.0001),但定量DNA水平并未随时间下降。储存温度对感染滴度或DNA水平均无显著影响。

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