Vanegas Hernan, González Fredman, Reyes Yaoska, Centeno Edwing, Palacios Jayrintzina, Zepeda Omar, Hagbom Marie, Collins Matthew H, Coward R Matthew, Becker-Dreps Sylvia, Bowman Natalie, Bucardo Filemón
Department of Microbiology, Faculty of Medical Science, National Autonomous University of Nicaragua, León 21000, Nicaragua.
Division of Molecular Virology, Department of Clinical and Experimental Medicine, Linköping University, SE-581 83 Linköping, Sweden.
Viruses. 2021 Jan 21;13(2):152. doi: 10.3390/v13020152.
Zika virus (ZIKV) RNA has been found to remain in human semen for up to one year after infection, but the presence of Flavivirus antigens in the different compartments of semen has been largely unexplored. Following the introduction of ZIKV in Nicaragua (2016), a prospective study of patients with clinical symptoms consistent with ZIKV was conducted in León to investigate virus shedding in different fluids. ZIKV infection was confirmed in 16 male subjects (≥18 years of age) by RT-qPCR in either blood, saliva or urine. Of these, three provided semen samples at 7, 14, 21, 28, 60 and 180 days postsymptom onset (DPSO) for Flavivirus antigens and RNA studies. These cases were compared with 19 asymptomatic controls. Flavivirus antigens were examined by immunofluorescence (IF) using the 4G2 Mabs, and confocal microscopy was used to explore fluorescence patterns. The three (100%) symptomatic subjects and 3 (16%) of the 19 asymptomatic subjects had Flavivirus antigens and viral RNA in the spermatozoa fraction. The percentage of IF Flavivirus-positive spermatozoa cells ranged from 1.9% to 25% in specimens from symptomatic subjects, as compared with 0.8% to 3.8% in specimens from asymptomatic controls. A marked IF-pattern in the cytoplasmic droplets and tail of the spermatozoa was observed. The sperm concentrations (45 × 10/mL vs. 63.5 × 10/mL, = 0.041) and the total motility percentage (54% vs. 75%, = 0.009) were significantly lower in specimens from ZIKV-positive than in those of ZIKV-negative. In conclusion, this study demonstrated the presence of Flavivirus antigens and RNA within a time frame of 28 DPSO in sperm cells of symptomatic and asymptomatic subjects during the ZIKV epidemic. These findings have implications for public health, in terms of nonarthropod-born, silent transmission facilitated by sperm cells and potential transmission from asymptomatic males to pregnant women, with consequences to the fetus.
寨卡病毒（ZIKV）RNA在感染后可在人类精液中留存长达一年，但精液不同组分中黄病毒抗原的存在情况在很大程度上尚未得到研究。在寨卡病毒传入尼加拉瓜（2016年）后，在莱昂对有与寨卡病毒相符临床症状的患者开展了一项前瞻性研究，以调查不同体液中的病毒排泄情况。通过逆转录定量聚合酶链反应（RT-qPCR）在16名男性受试者（≥18岁）的血液、唾液或尿液中确认了寨卡病毒感染。其中，三人在症状出现后7、14、21、28、60和180天（DPSO）提供了精液样本，用于黄病毒抗原和RNA研究。将这些病例与19名无症状对照者进行比较。使用4G2单克隆抗体通过免疫荧光（IF）检测黄病毒抗原，并使用共聚焦显微镜探索荧光模式。三名（100%）有症状受试者和19名无症状受试者中的3名（16%）在精子部分检测到黄病毒抗原和病毒RNA。有症状受试者样本中IF黄病毒阳性精子细胞的百分比在1.9%至25%之间，而无症状对照者样本中的这一比例为0.8%至3.8%。在精子的细胞质滴和尾部观察到明显的IF模式。寨卡病毒阳性样本中的精子浓度（45×10/mL对63.5×10/mL，P = 0.041）和总活力百分比（54%对75%，P = 0.009）显著低于寨卡病毒阴性样本。总之，本研究证明在寨卡病毒流行期间，有症状和无症状受试者的精子细胞在症状出现后28天内存在黄病毒抗原和RNA。这些发现对公共卫生具有重要意义，涉及精子细胞促成的非节肢动物传播、无声传播以及无症状男性向孕妇的潜在传播及其对胎儿的影响。
