Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil.
Isolation of Infective Zika Virus from Urine and Saliva of Patients in Brazil.
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DOI：
10.1371/journal.pntd.0004816
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发表时间：
2016-06
影响因子：
3.8
通讯作者：
Brasil P
中科院分区：
文献类型：
--
作者：
Bonaldo MC;Ribeiro IP;Lima NS;Dos Santos AA;Menezes LS;da Cruz SO;de Mello IS;Furtado ND;de Moura EE;Damasceno L;da Silva KA;de Castro MG;Gerber AL;de Almeida LG;Lourenço-de-Oliveira R;Vasconcelos AT;Brasil P
寨卡病毒（ZIKV）是一种引发全球爆发性疫情的紧急威胁。自2015年1月以来，41个国家报告了本土病例。在巴西，吉兰-巴雷综合征和小头畸形病例的增加与ZIKV感染有关。最近的一份报告描述了其主要假定载体Ae的低实验传播效率。埃及人与明显的性传播通知一起，促使对病毒传播的其他潜在来源进行调查。尿液和唾液先前已被确定为ZIKV诊断中的有用工具。在这里，我们描述了在巴西里约热内卢州从急性期患者的唾液和尿液中存在和分离传染性ZIKV颗粒。来自里约热内卢的9名患者的9份尿液和5份唾液样本在Vero细胞培养物中接种，并分别通过NAT-Zika，RT-PCR和RT-qPCR进行特异性ZIKV RNA检测和定量。获得了两个ZIKV分离株，一个来自尿液，一个来自唾液样本。通过所有方法在四名患者中鉴定出ZIKV核酸。当尿液和唾液样品均来自同一患者时，尿液病毒载量更高，证实了它是ZIKV分子诊断的更好来源的一般意义。尽管如此，从两个分离的菌株中，每一个来自一个病人，只有一个来自尿液，这表明其他因素，如这种液体的酸性，可能会干扰病毒粒子的感染性。获得两种ZIKV分离株的完整基因组。系统发育分析显示，与以前在南美疫情期间分离的菌株相似。在急性期期间在患者的尿液和唾液中检测到感染性ZIKV颗粒可能代表病毒传播的关键因素。关于替代非媒介ZIKV传播途径的贡献，这一发现的流行病学相关性需要进一步调查。最近，美洲大陆出现了寨卡病毒及其严重表现的毁灭性流行，如新生儿小头畸形和格林-巴利综合征。寨卡病毒于1947年在非洲首次发现，仅从2007年开始引发疫情。寨卡病毒、登革热和基孔肯雅病毒主要由伊蚊传播。登革热在巴西流行已近30年，该国主要受到其主要媒介埃及伊蚊的侵扰。基孔肯雅病毒于2014年底进入该国，8个月后证实存在寨卡病毒。尽管如此，寨卡病毒的通知成倍增加，并以前所未有的速度在全国蔓延，增加了其他传播途径的可能性。最近一些关于寨卡病毒在埃岛性传播的报道加强了这一假设。寨卡病毒的传播效率很低。埃及人。我们在急性期患者的尿液和唾液中发现了寨卡病毒活性颗粒，Fiocruz在2016年2月5日的新闻发布会上迅速宣布了这一发现。在这项工作中，我们提出了潜在的替代人与人之间的感染途径以外的媒介传播，可能有流行病学的相关性。
Zika virus (ZIKV) is an emergent threat provoking a worldwide explosive outbreak. Since January 2015, 41 countries reported autochthonous cases. In Brazil, an increase in Guillain-Barré syndrome and microcephaly cases was linked to ZIKV infections. A recent report describing low experimental transmission efficiency of its main putative vector, Ae. aegypti, in conjunction with apparent sexual transmission notifications, prompted the investigation of other potential sources of viral dissemination. Urine and saliva have been previously established as useful tools in ZIKV diagnosis. Here, we described the presence and isolation of infectious ZIKV particles from saliva and urine of acute phase patients in the Rio de Janeiro state, Brazil. Nine urine and five saliva samples from nine patients from Rio de Janeiro presenting rash and other typical Zika acute phase symptoms were inoculated in Vero cell culture and submitted to specific ZIKV RNA detection and quantification through, respectively, NAT-Zika, RT-PCR and RT-qPCR. Two ZIKV isolates were achieved, one from urine and one from saliva specimens. ZIKV nucleic acid was identified by all methods in four patients. Whenever both urine and saliva samples were available from the same patient, urine viral loads were higher, corroborating the general sense that it is a better source for ZIKV molecular diagnostic. In spite of this, from the two isolated strains, each from one patient, only one derived from urine, suggesting that other factors, like the acidic nature of this fluid, might interfere with virion infectivity. The complete genome of both ZIKV isolates was obtained. Phylogenetic analysis revealed similarity with strains previously isolated during the South America outbreak. The detection of infectious ZIKV particles in urine and saliva of patients during the acute phase may represent a critical factor in the spread of virus. The epidemiological relevance of this finding, regarding the contribution of alternative non-vectorial ZIKV transmission routes, needs further investigation. The American continent has recently been the scene of a devastating epidemic of Zika virus and its severe manifestations, such as microcephaly in newborns and Guillain-Barré Syndrome. Zika virus, first detected in 1947 in Africa, only from 2007 started provoking outbreaks. Zika, dengue and chikungunya viruses are primarily transmitted by Aedes mosquitoes. Dengue has been endemic in Brazil for almost 30 years, and the country is largely infested by its main vector, Aedes aegypti. Chikungunya virus entered the country in late 2014 and Zika presence was confirmed eight months later. Nevertheless, Zika notifications multiplied and spread across the country with unprecedented speed, raising the possibility of other transmission routes. This hypothesis was strengthened by some recent reports of Zika sexual transmission in Ae. aegypti-free areas and by the description of a low transmission efficiency to Zika virus in local Ae. aegypti. We found Zika active particles in both urine and saliva of acute phase patients, and a finding that was promptly announced by Fiocruz via Press Conference on February 5, 2016. In this work, we bring up the potential alternative person-to-person infection routes beyond the vectorial transmission, that might have epidemiological relevance.