Development of Moore Swab and Ultrafiltration Concentration and Detection Methods for Salmonella Typhi and Salmonella Paratyphi A in Wastewater and Application in Kolkata, India and Dhaka, Bangladesh.
Development of Moore Swab and Ultrafiltration Concentration and Detection Methods for Salmonella Typhi and Salmonella Paratyphi A in Wastewater and Application in Kolkata, India and Dhaka, Bangladesh.
复制标题
DOI：
10.3389/fmicb.2021.684094
复制
发表时间：
2021
影响因子：
5.2
通讯作者：
Moe CL
中科院分区：
文献类型：
--
作者：
Liu P;Ibaraki M;Kapoor R;Amin N;Das A;Miah R;Mukhopadhyay AK;Rahman M;Dutta S;Moe CL
肠道发热是由伤寒沙门氏菌（ST）和甲型副伤寒沙门氏菌（SPA）引起的严重全身感染。废水中ST和SPA的检测可用作监测战略，以确定感染负担，并确定水、环境卫生和个人卫生干预措施以及疫苗接种运动的优先领域。然而，环境样品中ST和SPA的敏感和特异性检测一直具有挑战性。在这项研究中，我们开发并验证了两种方法浓缩和检测废水中的ST/SPA：摩尔拭子捕获法的定性结果，超滤（UF）的灵敏定量检测，再加上qPCR。然后，我们将这些方法应用于ST和SPA废水监测在加尔各答，印度和达卡，孟加拉国，两个肠热病流行区。qPCR检测ST的检测限为17个当量基因组拷贝（EGC），SPA的检测限为25个EGC，具有良好的重现性。在接种试验中，ST和SPA的摩尔拭子法检测限约为0.05-0.005 cfu/mL。在2019年9月至2020年3月从加尔各答三个泵站采集的53份摩尔拭子样本中，ST检出率为69.8%，SPA检出率为20.8%。用已知量的ST和SPA接种并通过UF方法浓缩的污水样品的分析，然后通过聚乙二醇沉淀和qPCR检测表明，UF可以有效地回收5、10和20 L废水中约8、5和3 log 10 cfu的接种ST和SPA。在达卡使用UF方法，在26.7%（8/30）的20 L排水样本中检测到ST，范围为0.11-2.10 log 10 EGC/100 mL，在100%（4/4）的20 L根管样本中检测到ST，范围为1.02-2.02 log 10 EGC/100 mL。这些结果表明，摩尔拭子和UF方法提供了灵敏的存在/不存在和定量检测废水样品中的ST/SPA。
Enteric fever is a severe systemic infection caused by Salmonella enterica serovar Typhi (ST) and Salmonella enterica serovar Paratyphi A (SPA). Detection of ST and SPA in wastewater can be used as a surveillance strategy to determine burden of infection and identify priority areas for water, sanitation, and hygiene interventions and vaccination campaigns. However, sensitive and specific detection of ST and SPA in environmental samples has been challenging. In this study, we developed and validated two methods for concentrating and detecting ST/SPA from wastewater: the Moore swab trap method for qualitative results, and ultrafiltration (UF) for sensitive quantitative detection, coupled with qPCR. We then applied these methods for ST and SPA wastewater surveillance in Kolkata, India and Dhaka, Bangladesh, two enteric fever endemic areas. The qPCR assays had a limit of detection of 17 equivalent genome copies (EGC) for ST and 25 EGC for SPA with good reproducibility. In seeded trials, the Moore swab method had a limit of detection of approximately 0.05–0.005 cfu/mL for both ST and SPA. In 53 Moore swab samples collected from three Kolkata pumping stations between September 2019 and March 2020, ST was detected in 69.8% and SPA was detected in 20.8%. Analysis of sewage samples seeded with known amount of ST and SPA and concentrated via the UF method, followed by polyethylene glycol precipitation and qPCR detection demonstrated that UF can effectively recover approximately 8, 5, and 3 log10 cfu of seeded ST and SPA in 5, 10, and 20 L of wastewater. Using the UF method in Dhaka, ST was detected in 26.7% (8/30) of 20 L drain samples with a range of 0.11–2.10 log10 EGC per 100 mL and 100% (4/4) of 20 L canal samples with a range of 1.02–2.02 log10 EGC per 100 mL. These results indicate that the Moore swab and UF methods provide sensitive presence/absence and quantitative detection of ST/SPA in wastewater samples.
登录
查看更多内容
影响因子：
9.4
作者：
Manor, Y;Handsher, R;Mendelson, E
通讯作者：
Mendelson, E
DOI：
10.1016/j.ijheh.2020.113669
发表时间：
2021-02-09
影响因子：
6
作者：
Foster, Tim;Falletta, Jay;Willetts, Juliet
通讯作者：
Willetts, Juliet
影响因子：
11.8
作者：
Crump, John A.
通讯作者：
Crump, John A.
DOI：
10.1017/s0022172400028230
发表时间：
1968-01-01
期刊：
JOURNAL OF HYGIENE-CAMBRIDGE
影响因子：
--
作者：
CALLAGHAN, P;BRODIE, J
通讯作者：
BRODIE, J
DOI：
10.1017/s0022172400022300
发表时间：
1972-01-01
期刊：
JOURNAL OF HYGIENE-CAMBRIDGE
影响因子：
--
作者：
CONN, NK;SCOTT, TG;MCWILLIAM, JM
通讯作者：
MCWILLIAM, JM