Fedorko Daniel P, Preuss Jeanne C, Fahle Gary A, Li Li, Fischer Steven H, Hohman Patricia, Cohen Jeffrey I
Warren G. Magnuson Clinical Center, National Institutes for Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892-1508, USA.
J Clin Microbiol. 2005 Sep;43(9):4602-6. doi: 10.1128/JCM.43.9.4602-4606.2005.
We analyzed a shell vial culture assay (SVA), real-time PCR, and a direct fluorescent antibody assay (DFA) for rapid detection of vaccinia virus from vaccination sites of Dryvax vaccine recipients. Of 47 samples assayed, 100% were positive by PCR, 89% were positive by SVA, and 40% were positive by DFA. DFA was limited by the need for adequate numbers of cells, with 32% of samples inadequate for interpretation. DFA performed better with specimens from patients who had not previously received the vaccine. PCR was positive for longer times postvaccination than was SVA. Infectious virus could be recovered after 45 min of acetone fixation of shell vial coverslips. Commercially available polyclonal antibodies cross-reacted with other orthopoxviruses and herpes simplex 1, but commercially available monoclonal antibodies were specific for vaccinia virus. In summary, PCR was the most sensitive test for detecting vaccinia virus in clinical specimens, while the DFA was the most rapid but the least sensitive test.
我们分析了用于从Dryvax疫苗接种者接种部位快速检测痘苗病毒的空斑小瓶培养试验（SVA）、实时聚合酶链反应（PCR）和直接荧光抗体试验（DFA）。在检测的47份样本中，PCR检测的阳性率为100%，SVA检测的阳性率为89%，DFA检测的阳性率为40%。DFA受限于需要足够数量的细胞，32%的样本因细胞数量不足而无法进行判读。对于未曾接种过该疫苗的患者的样本，DFA的表现更佳。接种疫苗后，PCR检测呈阳性的时间比SVA长。空斑小瓶盖玻片经丙酮固定45分钟后仍可回收感染性病毒。市售多克隆抗体与其他正痘病毒和单纯疱疹病毒1发生交叉反应，但市售单克隆抗体对痘苗病毒具有特异性。总之，PCR是检测临床标本中痘苗病毒最敏感的试验，而DFA是最快速但最不敏感的试验。
