High-throughput low-cost nl-qPCR for enteropathogen detection: A proof-of-concept among hospitalized patients in Bangladesh.
High-throughput low-cost nl-qPCR for enteropathogen detection: A proof-of-concept among hospitalized patients in Bangladesh.
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DOI：
10.1371/journal.pone.0257708
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发表时间：
2021
期刊：
影响因子：
3.7
通讯作者：
Nelson EJ
中科院分区：
文献类型：
--
作者：
Flaherty KE;Grembi JA;Ramachandran VV;Haque F;Khatun S;Rahman M;Maples S;Becker TK;Spormann AM;Schoolnik GK;Hryckowian AJ;Nelson EJ
腹泻病是全球发病和死亡的主要原因，特别是在低收入和中等收入国家。需要采用高通量和低成本的方法来识别病原体，以指导公共卫生缓解措施。纳升 qPCR (nl-qPCR) 是一种较昂贵方法的有吸引力的替代方案，但其应用仍处于萌芽阶段，并且尚未在住院患者中进行概念验证。一项基于人口普查的研究是在腹泻爆发期间孟加拉国农村地区两家政府医院收治的腹泻患者中进行的。从粪便样本中提取 DNA，并通过 nl-qPCR 检测常见细菌、原生动物和蠕虫肠道病原体作为主要结果。共有 961 名患者入组；收集了 827 名患者的粪便样本。 69% 的患者样本中检测到肠道病原体； 32% 的人检测到一种以上肠道病原体。最常见的肠道病原体是肠聚集性大肠杆菌（26.0%）、产志贺毒素大肠杆菌（18.3%）、产肠毒素大肠杆菌（15.5%热稳定毒素阳性、2.2%热不稳定毒素阳性）、志贺氏菌属。 (14.8%) 和霍乱弧菌 (9.0%)。地理空间分析显示，居住在距离研究医院最近的患者中，每位患者的病原体中位数和严重脱水病例的比例最高。”本研究证明了 nl-qPCR 作为住院患者肠道病原体检测的高通量低成本方法的概念验证。
Diarrheal disease is a leading cause of morbidity and mortality globally, especially in low- and middle-income countries. High-throughput and low-cost approaches to identify etiologic agents are needed to guide public health mitigation. Nanoliter-qPCR (nl-qPCR) is an attractive alternative to more expensive methods yet is nascent in application and without a proof-of-concept among hospitalized patients. A census-based study was conducted among diarrheal patients admitted at two government hospitals in rural Bangladesh during a diarrheal outbreak period. DNA was extracted from stool samples and assayed by nl-qPCR for common bacterial, protozoan, and helminth enteropathogens as the primary outcome. A total of 961 patients were enrolled; stool samples were collected from 827 patients. Enteropathogens were detected in 69% of patient samples; More than one enteropathogen was detected in 32%. Enteropathogens most commonly detected were enteroaggregative Escherichia coli (26.0%), Shiga toxin-producing E.coli (18.3%), enterotoxigenic E. coli (15.5% heat stable toxin positive, 2.2% heat labile toxin positive), Shigella spp. (14.8%), and Vibrio cholerae (9.0%). Geospatial analysis revealed that the median number of pathogens per patient and the proportion of cases presenting with severe dehydration were greatest amongst patients residing closest to the study hospitals.” This study demonstrates a proof-of-concept for nl-qPCR as a high-throughput low-cost method for enteropathogen detection among hospitalized patients.
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