Zika virus (ZIKV) infection remains a public health concern necessitating demand for long-term virus production for diagnostic assays and R&D activities. Inactivated virus constitutes an important component of the Trioplex rRT-PCR assay and serological IgM assay (MAC-ELISA). The aim of our study is to establish standard methods of ZIKV inactivation while maintaining antigenicity and RNA integrity. We tested viral supernatants by four different inactivation methods: 1. Heat inactivation at 56 °C and 60 °C; 2. Gamma-Irradiation; 3. Chemical inactivation by Beta-propiolactone (BPL) and 4. Fast-acting commercial disinfecting agents. Effectivity was measured by cytopathic effect (CPE) and plaque assay. RNA stability and antigenicity were measured by RT-PCR and MAC-ELISA, respectively. Results: Heat inactivation: Low titer samples, incubated at 56 °C for 2 h, showed neither CPE or plaques compared to high titer supernatants that required 2.5 h. Inactivation occurred at 60 °C for 60 min with all virus titers. Gamma irradiation: Samples irradiated at ≥3 Mrad for low virus concentrations and ≥5Mrad for high virus titer completely inactivated virus. Chemical Inactivation: Neither CPE nor plaques were observed with ≥0.045 % BPL inactivation of ZIKV. Disinfectant: Treatment of viral supernatants with Micro-Chem Plus™, inactivated virus in 2 min, whereas, Ethanol (70 %) and STERIS Coverage® Spray TB inactivated the virus in 5 min.

寨卡病毒 (ZIKV) 感染仍然是一个公共卫生问题，因此需要长期生产病毒以用于诊断分析和研发活动。灭活病毒是 Trioplex rRT-PCR 测定和血清学 IgM 测定 (MAC-ELISA) 的重要组成部分。我们研究的目的是建立 ZIKV 灭活的标准方法，同时保持抗原性和 RNA 完整性。我们通过四种不同的灭活方法测试了病毒上清液： 1. 56℃和60℃热灭活； 2. 伽马射线照射； 3. β-丙内酯 (BPL) 化学灭活和 4. 速效商业消毒剂。通过细胞病变效应（CPE）和噬菌斑测定来测量有效性。分别通过RT-PCR和MAC-ELISA测量RNA稳定性和抗原性。结果：热灭活：与需要 2.5 小时的高滴度上清液相比，在 56 °C 下孵育 2 小时的低滴度样品既没有显示出 CPE 也没有出现噬菌斑。所有病毒滴度均在 60°C 60 分钟内灭活。伽马射线照射：对于低病毒浓度，样品照射≥3Mrad；对于高病毒滴度，样品照射≥5Mrad，完全灭活病毒。化学灭活：ZIKV BPL 灭活率≥0.045% 时，未观察到 CPE 和斑块。消毒剂：用 Micro-Chem Plus™ 处理病毒上清液，2 分钟内灭活病毒，而乙醇 (70%) 和 STERIS Coverage® Spray TB 5 分钟内灭活病毒。