The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019.
The presence of SARS-CoV-2 RNA in human sewage in Santa Catarina, Brazil, November 2019.
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DOI：
10.1016/j.scitotenv.2021.146198
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发表时间：
2021-07-15
期刊：
影响因子：
--
通讯作者：
Rodríguez-Lázaro D
中科院分区：
文献类型：
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作者：
Fongaro G;Stoco PH;Souza DSM;Grisard EC;Magri ME;Rogovski P;Schörner MA;Barazzetti FH;Christoff AP;de Oliveira LFV;Bazzo ML;Wagner G;Hernández M;Rodríguez-Lázaro D
从2019年10月至2020年3月，对弗洛里亚诺波利斯（巴西圣卡塔里纳）的人类污水进行了严重急性呼吸综合征冠状病毒2（SARS-CoV 2）分析。将25 ml污水样品澄清，并使用甘氨酸缓冲液法结合聚乙二醇沉淀浓缩病毒，并使用商业试剂盒提取病毒RNA。采用RT-qPCR方法检测SARS冠状病毒2型RNA，采用靶向N1区、S区和两个RdRp区的寡核苷酸。所有阳性样本的结果均在独立实验室中通过不同的RT-qPCR系统进一步确认。对S和RdRp扩增子进行测序，以确认与SARS-CoV-2的同一性。使用两种策略进行基因组测序;序列独立的单引物扩增（SISPA）方法和使用Illumina的NGS的直接宏基因组学。2019年11月27日检测到SARS-CoV-2 RNA（5.49 ± 0.02 log 10 SARS-CoV-2基因组拷贝（GC）L−1），检测结果由独立实验室和基因组测序分析证实。随后3起事件中的样本经所有RT-qPCR检测均为阳性;这些阳性结果也得到了独立实验室的确认。平均负荷为5.83 ± 0.12 log 10 SARS-CoV-2 GC L−1，范围为5.49 ± 0.02 log 10 GC L−1（2019年11月27日）至6.68 ± 0.02 log 10 GC L−1（2020年3月4日）。我们的研究结果表明，自2019年11月以来，SARS-CoV-2可能在巴西社区中未被发现，早于美洲报告的首例病例（2020年1月21日）。
Human sewage from Florianopolis (Santa Catarina, Brazil) was analyzed for severe acute respiratory syndrome coronavirus-2 (SARS-CoV2) from October 2019 until March 2020. Twenty five ml of sewage samples were clarified and viruses concentrated using a glycine buffer method coupled with polyethylene glycol precipitation, and viral RNA extracted using a commercial kit. SARS-CoV-2 RNA was detected by RT-qPCR using oligonucleotides targeting N1, S and two RdRp regions. The results of all positive samples were further confirmed by a different RT-qPCR system in an independent laboratory. S and RdRp amplicons were sequenced to confirm identity with SARS-CoV-2. Genome sequencing was performed using two strategies; a sequence-independent single-primer amplification (SISPA) approach, and by direct metagenomics using Illumina's NGS. SARS-CoV-2 RNA was detected on 27th November 2019 (5.49 ± 0.02 log10 SARS-CoV-2 genome copies (GC) L−1), detection being confirmed by an independent laboratory and genome sequencing analysis. The samples in the subsequent three events were positive by all RT-qPCR assays; these positive results were also confirmed by an independent laboratory. The average load was 5.83 ± 0.12 log10 SARS-CoV-2 GC L−1, ranging from 5.49 ± 0.02 log10 GC L−1 (27th November 2019) to 6.68 ± 0.02 log10 GC L−1 (4th March 2020). Our findings demonstrate that SARS-CoV-2 was likely circulating undetected in the community in Brazil since November 2019, earlier than the first reported case in the Americas (21st January 2020).
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影响因子：
48
作者：
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通讯作者：
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影响因子：
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作者：
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作者：
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DOI：
10.1590/s0074-02762009000400007
发表时间：
2009-07-01
期刊：
Memórias do Instituto Oswaldo Cruz
影响因子：
--
作者：
Schlindwein, AD;Simões, CMO;Barardi, CRM
通讯作者：
Barardi, CRM