Surveillance of Shiga Toxin–Producing <i>Escherichia coli</i> in Beef with Effective Procedures, Independent of Serotype

Hara-Kudo, Yukiko; Niizuma, Jun; Goto, I; Iizuka, Shinji; Kaji, Yoshifumi; Kamakura, Kazumasa; Suzuki, Sosuke; Takatori, Kosuke

doi:10.1089/fpd.2007.0031

Cited by 9 publications

(5 citation statements)

References 21 publications

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“…Previously, spiked samples were usually enriched overnight without characterizing the effects of different enrichment times on the detection outcomes [23], [24]. In this study, P. shigelloides strain ATCC 51903 was used in experiments with simulated human stool samples, with the LAMP assays having a detection limit of 5×10 3 CFU/g stool.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Sensitive Detection of Plesiomonas shigelloides by Loop-Mediated Isothermal Amplification of the hugA Gene

Shao¹,

Xu²,

Xiong³

et al. 2012

PLoS ONE

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Plesiomonas shigelloides is one of the causative agents of human gastroenteritis, with increasing number of reports describing such infections in recent years. In this study, the hugA gene was chosen as the target to design loop-mediated isothermal amplification (LAMP) assays for the rapid, specific, and sensitive detection of P. shigelloides. The performance of the assay with reference plasmids and spiked human stools as samples was evaluated and compared with those of quantitative PCR (qPCR). No false-positive results were observed for the 32 non-P. shigelloides strains used to evaluate assay specificity. The limit of detection for P. shigelloides was approximately 20 copies per reaction in reference plasmids and 5×103 CFU per gram in spiked human stool, which were more sensitive than the results of qPCR. When applied in human stool samples spiked with 2 low levels of P. shigelloides, the LAMP assays achieved accurate detection after 6-h enrichment. In conclusion, the LAMP assay developed in this study is a valuable method for rapid, cost-effective, and simple detection of P. shigelloides in basic clinical and field laboratories in the rural areas of China.

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Section: Discussionmentioning

confidence: 99%

Rapid and Sensitive Detection of Plesiomonas shigelloides by Loop-Mediated Isothermal Amplification of the hugA Gene

Shao¹,

Xu²,

Xiong³

et al. 2012

PLoS ONE

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“…Since it is isothermal, LAMP can be performed in much simpler instruments, such as a heater or water bath. To date, multiple LAMP assays targeting STEC Shiga toxin genes (stx 1 and stx 2 ) have been developed (20)(21)(22)(23)(24) and evaluated in food, primarily beef (25)(26)(27)(28), as have several others targeting the E. coli O157 rfbE gene (encoding perosamine synthetase) (23,24,(29)(30)(31). Very recently, we developed a suite of LAMP assays for STEC (targeting common virulence genes stx 1 , stx 2 , and eae) and the seven adulterant STEC serogroups (targeting the wzx or wzy gene on the respective O-antigen gene clusters) (32,33).…”

mentioning

confidence: 99%

Evaluation of a Loop-Mediated Isothermal Amplification Suite for the Rapid, Reliable, and Robust Detection of Shiga Toxin-Producing Escherichia coli in Produce

Wang

Yang

et al. 2014

Appl Environ Microbiol

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c Shiga toxin-producing Escherichia coli (STEC) strains are a leading cause of produce-associated outbreaks in the United States. Rapid, reliable, and robust detection methods are needed to better ensure produce safety. We recently developed a loop-mediated isothermal amplification (LAMP) suite for STEC detection. In this study, the STEC LAMP suite was comprehensively evaluated against real-time quantitative PCR (qPCR) using a large panel of bacterial strains (n ‫؍‬ 156) and various produce items (several varieties of lettuce, spinach, and sprouts). To simulate real-world contamination events, produce samples were surface inoculated with a low level (1.2 to 1.8 CFU/25 g) of individual STEC strains belonging to seven serogroups (O26, O45, O103, O111, O121, O145, and O157) and held at 4°C for 48 h before testing. Six DNA extraction methods were also compared using produce enrichment broths. All STEC targets and their subtypes were accurately detected by the LAMP suite. The detection limits were 1 to 20 cells per reaction in pure culture and 10 5 to 10 6 CFU per 25 g (i.e., 10 3 to 10 4 CFU per g) in produce, except for strains harboring the stx 2c , eae-␤, and eae-subtypes. After 6 to 8 h of enrichment, the LAMP suite achieved accurate detection of low levels of STEC strains of various stx 2 and eae subtypes in lettuce and spinach varieties but not in sprouts. A similar trend of detection was observed for qPCR. The PrepMan Ultra sample preparation reagent yielded the best results among the six DNA extraction methods. This research provided a rapid, reliable, and robust method for detecting STEC in produce during routine sampling and testing. The challenge with sprouts detection by both LAMP and qPCR calls for special attention to further analysis.

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“…Studies were also conducted to detect the serotypes of Escherichia coli (E. coli) O157, O26, and O111, which cause enterohemorrhagic infections, and the serotypes of E. coli O26, O45, O103, O111, O121, and O145, which produce Shiga toxin. [86][87][88]…”

Section: Lamp For Foodborne Illnesses Diagnosismentioning

confidence: 99%

Point‐of‐Care Diagnostic Platforms for Loop‐Mediated Isothermal Amplification

et al. 2022

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The loop‐mediated isothermal amplification (LAMP) method is one of the Nucleic acid amplification tests (NAATs) that allows for the amplification of target regions without using a thermal cycle. With its unique primer design, LAMP ensures the rapid replication of the targeted DNA region with high specificity and high efficiency. LAMP technology is used for diagnostic purposes in pathogen detection due to its ease of use, low cost, and simplicity without requiring complex equipment. A wide range of LAMP diagnostic platforms have been developed for applications in bacteria, virus, and parasitic pathogen detection. Herein, the methodology of LAMP technology and its applications in pathogen detection and SNP genotyping and mutation detection are discussed. Point‐of‐care (PoC) LAMP platforms designed with the principles of microfluidic chip technology, including LAMP‐on‐a‐chip, paper‐based LAMP, and smartphone‐based LAMP applications have been elaborated. LAMP technology represents a fast, robust, and reliable diagnostic platform for point‐of‐care testing.

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Surveillance of Shiga Toxin–Producing Escherichia coli in Beef with Effective Procedures, Independent of Serotype

Cited by 9 publications

References 21 publications

Rapid and Sensitive Detection of Plesiomonas shigelloides by Loop-Mediated Isothermal Amplification of the hugA Gene

Rapid and Sensitive Detection of Plesiomonas shigelloides by Loop-Mediated Isothermal Amplification of the hugA Gene

Evaluation of a Loop-Mediated Isothermal Amplification Suite for the Rapid, Reliable, and Robust Detection of Shiga Toxin-Producing Escherichia coli in Produce

Point‐of‐Care Diagnostic Platforms for Loop‐Mediated Isothermal Amplification

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