Institute of Virology and Antiviral Therapy, German Reference Laboratory for HSV and VZV, Jena University Clinic, Friedrich Schiller University Jena, Jena, Hans-Knoell-Strasse 2, 07745 Jena, Germany.
J Virol Methods. 2013 Mar;188(1-2):70-2. doi: 10.1016/j.jviromet.2012.12.001. Epub 2012 Dec 12.
Rapid alternative methods are required to evaluate easily acyclovir (ACV) sensitivity of clinical herpes simplex virus (HSV) isolates. The objective of this study was to screen 54 ACV-sensitive and 41 ACV-resistant clinical HSV-1 isolates, well characterized by phenotypic and genotypic methods, for the phosphorylation activity of the viral thymidine kinase (TK) using a commercially available and modified non-radioactive DiviTum® test on the basis of an indirect enzyme linked immunosorbent assay. The ACV-sensitive HSV-1 isolates had high TK activity values between 31.5±6.4 DiviTum® Units per liter (DU/L) and 487.4±60.1 DU/L. The mean activity of all ACV-sensitive isolates was calculated as 212.3±15.7 DU/L. By contrast, the mean activity of all ACV-resistant HSV-1 isolates was significantly lower at 5.5±1.3 DU/L. Out of the 41 ACV-resistant HSV-1 isolates, 38 had no or very low phosphorylation activities of the viral TK between 0 DU/L and 9.3±3.2 DU/L. The remaining three ACV-resistant viral isolates had TK activities between 44.6±5.1 DU/L and 80.9±13.3D U/L. In conclusion, the modified DiviTum® test can be used to screen HSV-1 isolates for their sensitivity to ACV. Acyclovir-sensitive HSV-1 isolates show TK activities >30 DU/L and ACV-resistant isolates have activity values <10 DU/L. However, single ACV-resistant HSV-1 isolates can have TK activity values >30 DU/L. These strains are most likely ACV-resistant TK-altered mutants, but no evidence was provided for an alteration of the TK.
需要快速替代方法来评估临床单纯疱疹病毒 (HSV) 分离物对阿昔洛韦 (ACV) 的敏感性。本研究的目的是使用市售且经过改良的非放射性 DiviTum®检测试剂盒，基于间接酶联免疫吸附试验，筛选 54 株 ACV 敏感和 41 株 ACV 耐药的临床 HSV-1 分离株的病毒胸苷激酶 (TK) 的磷酸化活性，这些分离株通过表型和基因型方法进行了很好的特征描述。ACV 敏感的 HSV-1 分离株的 TK 活性值在 31.5±6.4 DiviTum®单位/升 (DU/L) 到 487.4±60.1 DU/L 之间。所有 ACV 敏感分离株的平均活性计算为 212.3±15.7 DU/L。相比之下，所有 ACV 耐药 HSV-1 分离株的平均活性明显较低，为 5.5±1.3 DU/L。在 41 株 ACV 耐药的 HSV-1 分离株中，有 38 株的病毒 TK 磷酸化活性为 0 DU/L 或非常低，为 0 DU/L 至 9.3±3.2 DU/L。其余三个 ACV 耐药的病毒分离株的 TK 活性为 44.6±5.1 DU/L 和 80.9±13.3 DU/L。总之，改良的 DiviTum®检测可以用于筛选 HSV-1 分离物对 ACV 的敏感性。ACV 敏感的 HSV-1 分离株的 TK 活性>30 DU/L，而 ACV 耐药的分离株的活性值<10 DU/L。然而，单个 ACV 耐药的 HSV-1 分离株可能具有>30 DU/L 的 TK 活性。这些菌株最有可能是 ACV 耐药的 TK 改变突变体，但没有提供 TK 改变的证据。
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