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通过快速24孔板离心法以及使用地塞米松的传统细胞培养法检测腺病毒。

Detection of adenovirus by rapid 24-well plate centrifugation and conventional cell culture with dexamethasone.

作者信息

Woods G L, Yamamoto M, Young A

机构信息

Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha 68105.

出版信息

J Virol Methods. 1988 Jun;20(2):109-14. doi: 10.1016/0166-0934(88)90144-9.

Abstract

Two methods for rapid detection of adenovirus were tested: (i) 24-well plate centrifugation followed by staining with a monoclonal antibody after incubation for 24 h and 48 h, and (ii) pretreatment of A549 cells used in conventional cell culture and 24-well plate centrifugation with 10(-5)M dexamethasone. Twenty-seven clinical isolates of adenovirus and 12 specimens from which adenovirus had been recovered were included in the analysis. Both isolates and specimens had been frozen at -70 degrees C for up to 6 months. By 24-well plate centrifugation both with and without dexamethasone, 21 (78%) and 27 (100%) isolates were positive for adenovirus at 24 h and 48 h, respectively. Of the specimens, 6 (50%) and 8 (67%) were positive by 24-well plate centrifugation without dexamethasone at 24 h and 48 h, respectively, whereas with dexamethasone 3 (25%) were positive at 24 h and 7 (58%) were positive at 48 h. Overall, combining isolates and specimens, the sensitivity of 24-well plate centrifugation for detection of adenovirus at 24 h was 69% without dexamethasone and 62% with dexamethasone, and at 48 h the sensitivity was 90% without dexamethasone and 87% with dexamethasone. The specificity under all conditions tested was 100%. In conventional tissue culture dexamethasone inhibited recovery of adenovirus. Without dexamethasone, adenovirus was recovered from all 39 samples within 7 days after inoculation; however with dexamethasone pretreatment, the virus was detected in only 31 (79%) of the samples tested in the same period of time.

摘要

对两种快速检测腺病毒的方法进行了测试

(i)24孔板离心,在孵育24小时和48小时后用单克隆抗体染色;(ii)用10(-5)M地塞米松对常规细胞培养和24孔板离心所用的A549细胞进行预处理。分析纳入了27株腺病毒临床分离株和12份已从中分离出腺病毒的标本。分离株和标本均已在-70℃下冷冻长达6个月。通过24孔板离心,无论有无地塞米松,分别有21株(78%)和27株(100%)分离株在24小时和48小时时腺病毒检测呈阳性。在标本中,分别有6份(50%)和8份(67%)在24小时和48小时时通过无地塞米松的24孔板离心检测呈阳性,而使用地塞米松时,24小时时有3份(25%)呈阳性,48小时时有7份(58%)呈阳性。总体而言,将分离株和标本合并计算,无地塞米松时24孔板离心检测腺病毒在24小时时的灵敏度为69%,使用地塞米松时为62%,48小时时无地塞米松时灵敏度为90%,使用地塞米松时为87%。在所有测试条件下特异性均为100%。在常规组织培养中,地塞米松抑制腺病毒的回收。无地塞米松时,接种后7天内从所有39个样本中均回收了腺病毒;然而,经过地塞米松预处理后,在同一时间段内仅在31份(79%)测试样本中检测到该病毒。

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