PDF(Pubmed)
Abstract：
BACKGROUND: Rotavirus-induced viral gastroenteritis outbreaks result in over two million hospitalizations globally yearly. Wastewater-based epidemiology (WBE) has emerged as a crucial tool for detecting and monitoring viral outbreaks. The adoption of WBE has been instrumental in the early detection and surveillance of such viral outbreaks, providing a non-invasive method to assess public health.
OBJECTIVE: This study aims to utilize droplet digital polymerase chain reaction (ddPCR) technology to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India, thereby contributing to the understanding and management of viral gastroenteritis outbreaks through environmental surveillance.
METHODS: In this study, we used ddPCR to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India. We monitored its viral presence in municipal sewage treatment plants bi-weekly using an advanced ddPCR assay. Targeting the rotavirus non-structural protein 3 (NSP-3) region with custom primers and TaqMan probes, we detected virus concentration employing polyethylene glycol (PEG). Following RNA isolation, complementary DNA (cDNA) synthesis, and ddPCR analysis, our novel method eliminated standard curve dependence, propelling virus research and treatment forward.
RESULTS: Out of the 42 samples collected, a 16.60% positivity rate was observed, indicating a moderate presence of Rotavirus in Bhopal. The wastewater treatment plants (WWTP) attached to a hospital exhibited a 42.85% positivity rate, indicating the need for targeted monitoring. Leveraging ddPCR, precise quantification of rotavirus concentrations (ranging from 0.75 to 28.9 copies/µL) facilitated understanding and supported effective remediation.
CONCLUSIONS: This study emphasizes the importance of vigilant wastewater surveillance, especially in WWTPs with higher rotavirus prevalence. The significance of ddPCR in comparison to conventional and real-time PCR lies in its superior sensitivity and specificity in detecting and quantifying positive samples. Furthermore, it can identify positive samples even in the smallest quantities without the need for a standard curve to evaluate. This makes ddPCR a valuable tool for accurate and precise detection and quantification of samples.
OBJECTIVE: This study aims to utilize droplet digital polymerase chain reaction (ddPCR) technology to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India, thereby contributing to the understanding and management of viral gastroenteritis outbreaks through environmental surveillance.
METHODS: In this study, we used ddPCR to detect and quantify Rotavirus in wastewater samples collected from the Bhopal region of India. We monitored its viral presence in municipal sewage treatment plants bi-weekly using an advanced ddPCR assay. Targeting the rotavirus non-structural protein 3 (NSP-3) region with custom primers and TaqMan probes, we detected virus concentration employing polyethylene glycol (PEG). Following RNA isolation, complementary DNA (cDNA) synthesis, and ddPCR analysis, our novel method eliminated standard curve dependence, propelling virus research and treatment forward.
RESULTS: Out of the 42 samples collected, a 16.60% positivity rate was observed, indicating a moderate presence of Rotavirus in Bhopal. The wastewater treatment plants (WWTP) attached to a hospital exhibited a 42.85% positivity rate, indicating the need for targeted monitoring. Leveraging ddPCR, precise quantification of rotavirus concentrations (ranging from 0.75 to 28.9 copies/µL) facilitated understanding and supported effective remediation.
CONCLUSIONS: This study emphasizes the importance of vigilant wastewater surveillance, especially in WWTPs with higher rotavirus prevalence. The significance of ddPCR in comparison to conventional and real-time PCR lies in its superior sensitivity and specificity in detecting and quantifying positive samples. Furthermore, it can identify positive samples even in the smallest quantities without the need for a standard curve to evaluate. This makes ddPCR a valuable tool for accurate and precise detection and quantification of samples.
摘要：
背景：轮状病毒引起的病毒性胃肠炎暴发每年导致全球超过两百万人住院。基于废水的流行病学（WBE）已成为检测和监测病毒暴发的重要工具。WBE的采用有助于早期发现和监测此类病毒爆发，提供一种非侵入性的方法来评估公众健康。
目的：本研究旨在利用液滴数字聚合酶链反应（ddPCR）技术对从印度博帕尔地区收集的废水样品中的轮状病毒进行检测和定量，从而通过环境监测有助于了解和管理病毒性肠胃炎的爆发。
方法：在本研究中，我们使用ddPCR检测和定量从印度博帕尔地区收集的废水样品中的轮状病毒。我们使用先进的ddPCR测定法每两周监测其在市政污水处理厂中的病毒存在。用自定义引物和TaqMan探针靶向轮状病毒非结构蛋白3（NSP-3）区域，我们使用聚乙二醇（PEG）检测病毒浓度。RNA分离后,互补DNA（cDNA）合成，和ddPCR分析，我们的新方法消除了标准曲线依赖性，推动病毒研究和治疗向前发展。
结果：在收集的42个样本中，观察到16.60%的阳性率，表明轮状病毒在博帕尔的中度存在。附属于医院的污水处理厂（WWTP）的阳性率为42.85％，表明需要有针对性的监测。利用ddPCR，轮状病毒浓度的精确定量(范围为0.75至28.9拷贝/微升)有助于理解并支持有效补救.
结论：这项研究强调了警惕废水监测的重要性，特别是在轮状病毒流行率较高的WWTP中。与常规和实时PCR相比，ddPCR的重要性在于其在检测和定量阳性样品方面具有出色的灵敏度和特异性。此外，它可以识别阳性样品，即使在最小的数量，而不需要一个标准曲线来评估。这使得ddPCR成为准确和精确检测和定量样品的有价值的工具。
目的：本研究旨在利用液滴数字聚合酶链反应（ddPCR）技术对从印度博帕尔地区收集的废水样品中的轮状病毒进行检测和定量，从而通过环境监测有助于了解和管理病毒性肠胃炎的爆发。
方法：在本研究中，我们使用ddPCR检测和定量从印度博帕尔地区收集的废水样品中的轮状病毒。我们使用先进的ddPCR测定法每两周监测其在市政污水处理厂中的病毒存在。用自定义引物和TaqMan探针靶向轮状病毒非结构蛋白3（NSP-3）区域，我们使用聚乙二醇（PEG）检测病毒浓度。RNA分离后,互补DNA（cDNA）合成，和ddPCR分析，我们的新方法消除了标准曲线依赖性，推动病毒研究和治疗向前发展。
结果：在收集的42个样本中，观察到16.60%的阳性率，表明轮状病毒在博帕尔的中度存在。附属于医院的污水处理厂（WWTP）的阳性率为42.85％，表明需要有针对性的监测。利用ddPCR，轮状病毒浓度的精确定量(范围为0.75至28.9拷贝/微升)有助于理解并支持有效补救.
结论：这项研究强调了警惕废水监测的重要性，特别是在轮状病毒流行率较高的WWTP中。与常规和实时PCR相比，ddPCR的重要性在于其在检测和定量阳性样品方面具有出色的灵敏度和特异性。此外，它可以识别阳性样品，即使在最小的数量，而不需要一个标准曲线来评估。这使得ddPCR成为准确和精确检测和定量样品的有价值的工具。
