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
| Autor: | Mahbubur Rahman, Makoto Ibaraki, Abhishek Das, Pengbo Liu, Asish K. Mukhopadhyay, Shanta Dutta, Rana Miah, Nuhu Amin, Christine L. Moe, Renuka Kapoor |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Microbiology (medical) Veterinary medicine Salmonella enterica serovar Paratyphi A 030106 microbiology 030231 tropical medicine Ultrafiltration Sewage Biology Salmonella typhi Microbiology Salmonella Typhi 03 medical and health sciences 0302 clinical medicine wastewater Original Research business.industry Salmonella paratyphi A QR1-502 Wastewater Salmonella enterica serovar Typhi Moore swab Polyethylene glycol precipitation ultrafiltration surveillance business Salmonella Paratyphi A |
| Zdroj: | Frontiers in Microbiology Frontiers in Microbiology, Vol 12 (2021) |
| ISSN: | 1664-302X |
| Popis: | 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 to 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 log10 cfu, 5 log10 cfu, and 3 log10 cfu of seeded ST and SPA in 5 L, 10 L, 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, and these two methods can be used jointly or separately for Salmonella Typhi environmental surveillance. |
| Databáze: | OpenAIRE |
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