Evaluation of different target genes for the detection of
            <i>Salmonella</i>
            sp. by loop‐mediated isothermal amplification

Kreitlow, Antonia; Becker, André; Schotte, Ulrich; Malorny, Burkhard; Plötz, Madeleine; Abdulmawjood, Amir

doi:10.1111/lam.13409

Cited by 11 publications

(9 citation statements)

References 21 publications

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“…Previous studies have identified gene markers, such as fimA , hilA , invA , and ttr , that are useful in detecting Salmonella species ( Laing et al, 2017 ; Gand M. et al, 2020b ; Kreitlow et al, 2021 ). Moreover, for serovar identification, gene markers based on allelic variations in O and H antigen genes were used ( Laing et al, 2017 ; Gand M. et al, 2020b ).…”

Section: Introductionmentioning

confidence: 99%

Rapid Real-Time Polymerase Chain Reaction for Salmonella Serotyping Based on Novel Unique Gene Markers by Pangenome Analysis

Yang

Kim

et al. 2021

Front. Microbiol.

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An accurate diagnostic method for Salmonella serovars is fundamental to preventing the spread of associated diseases. A diagnostic polymerase chain reaction (PCR)-based method has proven to be an effective tool for detecting pathogenic bacteria. However, the gene markers currently used in real-time PCR to detect Salmonella serovars have low specificity and are developed for only a few serovars. Therefore, in this study, we explored the novel unique gene markers for 60 serovars that share similar antigenic formulas and show high prevalence using pangenome analysis and developed a real-time PCR to detect them. Before exploring gene markers, the 535 Salmonella genomes were evaluated, and some genomes had serovars different from the designated serovar information. Based on these analyses, serovar-specific gene markers were explored. These markers were identified as genes present in all strains of target serovar genomes but absent in strains of other serovar genomes. Serovar-specific primer pairs were designed from the gene markers, and a real-time PCR method that can distinguish between 60 of the most common Salmonella serovars in a single 96-well plate assay was developed. As a result, real-time PCR showed 100% specificity for 199 Salmonella and 29 non-Salmonella strains. Subsequently, the method developed was applied successfully to both strains with identified serovars and an unknown strain, demonstrating that real-time PCR can accurately detect serovars of strains compared with traditional serotyping methods, such as antisera agglutination. Therefore, our method enables rapid and economical Salmonella serotyping compared with the traditional serotyping method.

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Section: Introductionmentioning

confidence: 99%

Rapid Real-Time Polymerase Chain Reaction for Salmonella Serotyping Based on Novel Unique Gene Markers by Pangenome Analysis

Yang

Kim

et al. 2021

Front. Microbiol.

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“…The serovar-specific primers were selected for the criterium of exclusivity based on the in silico comparative genome analysis, but their target genes do not necessarily represent specific virulence genes or O and H antigen associated markers. The assay also includes a primer set for the type III secretion system-associated invA, a widely used gene target to confirm the genus of Salmonella [13,18]. All strains included in the study signaled positive using the selected invA primers (Table 3).…”

Section: Evaluation Using Characterized Salmonella Isolatesmentioning

confidence: 99%

“…First LAMP assays to identify Salmonella spp. and a few serovars in clinical and food samples have been described in recent years [10,[18][19][20].…”

Section: Introductionmentioning

confidence: 99%

Development of a rapid diagnostic test based on loop-mediated isothermal amplification to identify the most frequent non-typhoidal Salmonella serovars from culture

Edel

Glöckner

Stoll

et al. 2023

Eur J Clin Microbiol Infect Dis

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Identification of Salmonella serovars is performed by conventional seroagglutination or sequencing. These methods are labor-intensive and require technical experience. An easy-to-perform assay allowing the timely identification of the most common non-typhoidal serovars (NTS) is needed. In this study, a molecular assay based on loop-mediated isothermal amplification (LAMP) targeting specific gene sequences of Salmonella Enteritidis, S. Typhimurium, S. Infantis, S. Derby, and S. Choleraesuis has been developed for rapid serovar identification from cultured colonies. A total of 318 Salmonella strains and 25 isolates of other Enterobacterales species that served as negative controls were analyzed. All S. Enteritidis (n = 40), S. Infantis (n = 27), and S. Choleraesuis (n = 11) strains were correctly identified. Seven out of 104 S. Typhimurium and 10 out of 38 S. Derby strains missed a positive signal. Cross-reactions of the gene targets were only rarely observed and restricted to the S. Typhimurium primer set (5 false-positives). Sensitivity and specificity of the assay compared to seroagglutination were as follows: 100% and 100% for S. Enteritidis, 93.3% and 97.7% for S. Typhimurium, 100% and 100% for S. Infantis, 73.7% and 100% for S. Derby, and 100% and 100% for S. Choleraesuis, respectively. With results available in just a few minutes of hands-on time and a test run time of 20 min, the LAMP assay developed here may be a useful tool for the rapid identification of common Salmonella NTS in daily routine diagnostics.

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“…In Salmonella, the invA gene is an invasive protein gene, encoding a protein that establishes the invasive ability of Salmonella to intestinal mucosal cells. Besides, this gene is the most direct indicator of pathogenic Salmonella existence (Villamil et al 2020;Kreitlow et al 2021).…”

Section: Introductionmentioning

confidence: 99%

“…2020; Kreitlow et al . 2021). Another alternative gene for detecting Salmonella is the fimY gene, which has been detailed as one of a kind to the Salmonella species and regulates type 1 fimbrial expression (Tinker and Clegg 2000).…”

Section: Introductionmentioning

confidence: 99%

Sensitive and high-accuracy detection of Salmonella based on CRISPR/Cas12a combined with recombinase polymerase amplification

Mao¹,

Zhao²,

Jiang³

et al. 2022

Letters in Applied Microbiology

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Salmonella is a crucial food‐borne pathogen causing food poisoning, leading to severe public health events. Here, we developed a technique by integrating recombinase polymerase amplification with CRISPR‐LbCas12a and employing two targets with engineered crRNA for detection of Salmonella (RPA‐LbCas12a‐TTECDS). Our findings revealed that this novel method rapidly detects trace Salmonella in food through fluorescence intensity and provides a template for other food‐borne pathogen detection methods. Further, crRNA was optimized to increase detection sensitivity. Double targets were used to enhance the detection accuracy, reaching the level of qPCR, which was superior to fluorescent RPA. The RPA‐LbCas12a‐TTECDS system specifically detected Salmonella levels as low as 50 CFU per ml at 37°C in 1 h. In summary, a simple, rapid, sensitive and high accuracy detection technique based on CRISPR‐Cas12a was created for Salmonella detection without complicated equipment.

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Evaluation of different target genes for the detection of Salmonella sp. by loop‐mediated isothermal amplification

Cited by 11 publications

References 21 publications

Rapid Real-Time Polymerase Chain Reaction for Salmonella Serotyping Based on Novel Unique Gene Markers by Pangenome Analysis

Rapid Real-Time Polymerase Chain Reaction for Salmonella Serotyping Based on Novel Unique Gene Markers by Pangenome Analysis

Development of a rapid diagnostic test based on loop-mediated isothermal amplification to identify the most frequent non-typhoidal Salmonella serovars from culture

Sensitive and high-accuracy detection of Salmonella based on CRISPR/Cas12a combined with recombinase polymerase amplification

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