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北喀拉拉邦使用 Sanger 测序对 SARS-CoV-2 的 RBD 区域进行测序的基因组监测。

Genomic surveillance of SARS-CoV-2 by sequencing the RBD region using Sanger sequencing from North Kerala.

机构信息

Multidisciplinary Research Unit, Government Medical College, Kozhikode, Kerala, India.

Virus Research and Diagnostic Laboratory, Government Medical College, Kozhikode, Kerala, India.

出版信息

Front Public Health. 2022 Aug 25;10:974667. doi: 10.3389/fpubh.2022.974667. eCollection 2022.

Abstract

Next Generation Sequencing (NGS) is the gold standard for the detection of new variants of SARS-CoV-2 including those which have immune escape properties, high infectivity, and variable severity. This test is helpful in genomic surveillance, for planning appropriate and timely public health interventions. But labs with NGS facilities are not available in small or medium research settings due to the high cost of setting up such a facility. Transportation of samples from many places to few centers for NGS testing also produces delays due to transportation and sample overload leading in turn to delays in patient management and community interventions. This becomes more important for patients traveling from hotspot regions or those suspected of harboring a new variant. Another major issue is the high cost of NGS-based tests. Thus, it may not be a good option for an economically viable surveillance program requiring immediate result generation and patient follow-up. The current study used a cost-effective facility which can be set up in a common research lab and which is replicable in similar centers with expertise in Sanger nucleotide sequencing. More samples can be processed at a time and can generate the results in a maximum of 2 days (1 day for a 24 h working lab). We analyzed the nucleotide sequence of the Receptor Binding Domain (RBD) region of SARS-CoV-2 by the Sanger sequencing using in-house developed methods. The SARS-CoV-2 variant surveillance was done during the period of March 2021 to May 2022 in the Northern region of Kerala, a state in India with a population of 36.4 million, for implementing appropriate timely interventions. Our findings broadly agree with those from elsewhere in India and other countries during the period.

摘要

下一代测序(NGS)是检测 SARS-CoV-2 新变体的金标准,包括具有免疫逃逸特性、高传染性和可变严重程度的变体。该测试有助于进行基因组监测,为计划适当和及时的公共卫生干预措施提供参考。但是,由于建立此类设施的成本较高,因此小型或中型研究环境中没有配备 NGS 设施的实验室。由于运输和样本过载,将来自许多地方的样本运送到少数几个中心进行 NGS 测试也会导致延迟,从而导致患者管理和社区干预的延迟。对于来自热点地区的患者或那些疑似携带新变体的患者来说,这一点变得更加重要。另一个主要问题是 NGS 测试的成本很高。因此,对于需要立即生成结果并进行患者随访的经济可行的监测计划来说,这可能不是一个好的选择。本研究使用了一种具有成本效益的设施,可以在普通研究实验室中建立,并且在具有 Sanger 核苷酸测序专业知识的类似中心中具有可复制性。更多的样本可以同时处理,并在最多 2 天内(实验室工作 24 小时的情况下为 1 天)生成结果。我们使用内部开发的方法通过 Sanger 测序分析了 SARS-CoV-2 的受体结合域(RBD)区域的核苷酸序列。在印度喀拉拉邦北部地区,从 2021 年 3 月到 2022 年 5 月期间进行了 SARS-CoV-2 变体监测,以便及时实施适当的干预措施。我们的研究结果与印度其他地区和其他国家在同一时期的研究结果基本一致。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2b3c/9454329/70dc34708063/fpubh-10-974667-g0001.jpg

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