Detection of Salmonella enterica and Listeria monocytogenes in alternative irrigation water by culture and qPCR-based methods in the Mid-Atlantic U.S.
Detection of Salmonella enterica and Listeria monocytogenes in alternative irrigation water by culture and qPCR-based methods in the Mid-Atlantic U.S.
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DOI：
10.1128/spectrum.03536-23
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发表时间：
2024-02
影响因子：
3.7
通讯作者：
Chanelle L. Acheamfour;Salina Parveen;Alan Gutierrez;Eric T. Handy;Sara Behal;Donghyun Kim;Seongyun Kim;Cheryl L. East;Ray Xiong;Joseph Haymaker;S. Micallef;Rachel E. Rosenberg Goldstein;K. Kniel;Amy R. Sapkota;Fawzy Hashem;Manan Sharma
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文献类型：
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作者：
Chanelle L. Acheamfour;Salina Parveen;Alan Gutierrez;Eric T. Handy;Sara Behal;Donghyun Kim;Seongyun Kim;Cheryl L. East;Ray Xiong;Joseph Haymaker;S. Micallef;Rachel E. Rosenberg Goldstein;K. Kniel;Amy R. Sapkota;Fawzy Hashem;Manan Sharma
摘要替代灌溉沃茨（河流、池塘和再生水）可能含有细菌性食源性病原体，如肠道沙门氏菌和单核细胞增生李斯特菌，可能污染水果和蔬菜商品。与基于培养的方法相比，使用基于qPCR的方法检测食源性病原体可以加快检测方法和程序。本研究比较了S. enterica和L.通过qPCR（实时PCR）和培养方法在灌溉沃茨中检测单核细胞增生，以确定水类型（河流、池塘和再生水）、季节（冬季、春季、夏季和秋季）或体积（0.1、1和10 L）对这些方法的灵敏度、准确度、特异性和阳性（PPV）和阴性（NPV）预测值的影响。在美国大西洋中部的11个地点，通过改良摩尔拭子（MMS）过滤收集水样，为期2年，每两周或每月一次。对于qPCR，通过S特异性多重qPCR分析来自培养物富集样品（n = 1，990）的细菌DNA。enterica和L.单核细胞增多症对于培养物检测，选择性富集、分离富集样品，并进行PCR确认。PPV用于S. enterica和L.单核细胞增多症分别为68%和67%。净现值为87%（S. enterica）和85%（L.单核细胞增多症）。与秋季和冬季相比，春季和夏季观察到S. enterica; L.与单核细胞增生症相比，冬季的一致性水平低于春季、夏季和秋季。对于两种S. enterica和L.单核细胞增生，表明水的类型可能会影响这些结果的协议。检测灌溉水中的食源性病原体可以为干预和管理策略提供信息，以降低与新鲜和鲜切水果和蔬菜相关的污染和疾病风险。使用qPCR等非培养方法有可能加速检测过程。结果表明，池塘和再生水的文化和qPCR方法之间的协议比河水，可能是由于特定的理化特性的水。这些发现还表明季节和样品体积影响qPCR和培养结果的一致性。总体而言，qPCR方法可以更有把握地用于确定本研究中检查的灌溉水样品中是否存在肠道沙门氏菌和单核细胞增生李斯特菌。检测灌溉水中的食源性病原体可以为干预措施和管理策略提供信息，以减少与新鲜和鲜切水果和蔬菜相关的污染和疾病风险。使用qPCR等非培养方法有可能加速检测过程。结果表明，池塘和再生水的文化和qPCR方法之间的协议比河水，可能是由于特定的理化特性的水。这些发现还表明季节和样品体积影响qPCR和培养结果的一致性。总体而言，qPCR方法可以更有把握地用于确定本研究中检查的灌溉水样品中是否存在肠道沙门氏菌和单核细胞增生李斯特菌。
ABSTRACT Alternative irrigation waters (rivers, ponds, and reclaimed water) can harbor bacterial foodborne pathogens like Salmonella enterica and Listeria monocytogenes, potentially contaminating fruit and vegetable commodities. Detecting foodborne pathogens using qPCR-based methods may accelerate testing methods and procedures compared to culture-based methods. This study compared detection of S. enterica and L. monocytogenes by qPCR (real-time PCR) and culture methods in irrigation waters to determine the influence of water type (river, pond, and reclaimed water), season (winter, spring, summer, and fall), or volume (0.1, 1, and 10 L) on sensitivity, accuracy, specificity, and positive (PPV), and negative (NPV) predictive values of these methods. Water samples were collected by filtration through modified Moore swabs (MMS) over a 2-year period at 11 sites in the Mid-Atlantic U.S. on a bi-weekly or monthly schedule. For qPCR, bacterial DNA from culture-enriched samples (n = 1,990) was analyzed by multiplex qPCR specific for S. enterica and L. monocytogenes. For culture detection, enriched samples were selectively enriched, isolated, and PCR confirmed. PPVs for qPCR detection of S. enterica and L. monocytogenes were 68% and 67%, respectively. The NPV were 87% (S. enterica) and 85% (L. monocytogenes). Higher levels of qPCR/culture agreement were observed in spring and summer compared to fall and winter for S. enterica; for L. monocytogenes, lower levels of agreement were observed in winter compared to spring, summer, and fall. Reclaimed and pond water supported higher levels of qPCR/culture agreement compared to river water for both S. enterica and L. monocytogenes, indicating that water type may influence the agreement of these results. IMPORTANCE Detecting foodborne pathogens in irrigation water can inform interventions and management strategies to reduce risk of contamination and illness associated with fresh and fresh-cut fruits and vegetables. The use of non-culture methods like qPCR has the potential to accelerate the testing process. Results indicated that pond and reclaimed water showed higher levels of agreement between culture and qPCR methods than river water, perhaps due to specific physiochemical characteristics of the water. These findings also show that season and sample volume affect the agreement of qPCR and culture results. Overall, qPCR methods could be more confidently utilized to determine the absence of Salmonella enterica and Listeria monocytogenes in irrigation water samples examined in this study. Detecting foodborne pathogens in irrigation water can inform interventions and management strategies to reduce risk of contamination and illness associated with fresh and fresh-cut fruits and vegetables. The use of non-culture methods like qPCR has the potential to accelerate the testing process. Results indicated that pond and reclaimed water showed higher levels of agreement between culture and qPCR methods than river water, perhaps due to specific physiochemical characteristics of the water. These findings also show that season and sample volume affect the agreement of qPCR and culture results. Overall, qPCR methods could be more confidently utilized to determine the absence of Salmonella enterica and Listeria monocytogenes in irrigation water samples examined in this study.