Loop-mediated isothermal amplification (LAMP) for rapid detection of Salmonella in foods based on new molecular targets

Shang, Yuting; Ye, Qinghua; Cai, Shuzhen; Wu, Qingping; Pang, Rui; Yang, Shuanghong; Xiang, Xinran; Wang, Chu‐Fang; Zha, Fei; Ding, Yu; Zhang, Yinzhi; Wang, Jiasheng; Sun, Xiulan; Zhang, Jumei

doi:10.1016/j.lwt.2021.110999

Cited by 31 publications

(8 citation statements)

References 37 publications

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Section: Discussionmentioning

confidence: 99%

“…Molecular detection methods play an important role in rapidly detecting pathogenic bacteria [27]. The usefulness of molecular detection methods is dependent on the target genes or sequences and the specificity of specific primer pairs [27]. The current PCR methods for pathogenic staphylococci target 16S rRNA genes, housekeeping genes, or virulence genes [26,38].…”

Section: Discussionmentioning

confidence: 99%

“…However, some of these genes cannot discriminate between phylogenetically closely related species, such as the S. epidermidis and S. aureus cluster groups. The lack of exclusivity and inclusivity of molecular target genes is one major drawback of implementing the PCR-based method in food inspection [27]. Advances in high-throughput sequencing technology have allowed increasing the number of whole-genome sequences available in public databases, making it easier to obtain target genes specific for Staphylococcus species using bioinformatics approaches [27].…”

Section: Introductionmentioning

confidence: 99%

“…The lack of exclusivity and inclusivity of molecular target genes is one major drawback of implementing the PCR-based method in food inspection [27]. Advances in high-throughput sequencing technology have allowed increasing the number of whole-genome sequences available in public databases, making it easier to obtain target genes specific for Staphylococcus species using bioinformatics approaches [27]. This study aimed to develop a real-time PCR assay targeting molecular target genes to detect the staphylococci S. aureus, S. capitis, S. caprae, and S. epidermidis, thus allowing their accurate and rapid monitoring in food matrix.…”

Section: Introductionmentioning

confidence: 99%

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Real-Time PCR Method for the Rapid Detection and Quantification of Pathogenic Staphylococcus Species Based on Novel Molecular Target Genes

Kim

Yang

Won

et al. 2021

Foods

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Coagulase-positive Staphylococcus aureus is a foodborne pathogen considered one of the causes of food-related disease outbreaks. Like S. aureus, Staphylococcus capitis, Staphylococcus caprae, and S. epidermidis are opportunistic pathogens causing clinical infections and food contamination. The objective of our study was to develop a rapid, accurate, and monitoring technique to detect four Staphylococcus species in food. Four novel molecular targets (GntR family transcriptional regulator for S. aureus, phosphomannomutase for S. epidermidis, FAD-dependent urate hydroxylase for S. capitis, and Gram-positive signal peptide protein for S. caprae) were mined based on pan-genome analysis. Primers targeting molecular target genes showed 100% specificity for 100 non-target reference strains. The detection limit in pure cultures and artificially contaminated food samples was 102 colony-forming unit/mL for S. aureus, S. capitis, S. caprae, and S. epidermidis. Moreover, real-time polymerase chain reaction successfully detected strains isolated from various food matrices. Thus, our method allows an accurate and rapid monitoring of Staphylococcus species and may help control staphylococcal contamination of food.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Real-Time PCR Method for the Rapid Detection and Quantification of Pathogenic Staphylococcus Species Based on Novel Molecular Target Genes

Kim

Yang

Won

et al. 2021

Foods

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“…Nucleic acid detection methods have been widely used in clinical and in-field detection of microorganisms. 10 In the recent years, global outbreaks of human and animal epidemic, such as COVID-19 and Africa Swine Fever, have dramatically accelerated the technical development and practical applications of emerging nucleic acid detection methods, and various isothermal amplification technologies have been frequently reported for bacteria detection, including strand exchange amplification (SEA), 11 loop-mediated isothermal amplification (LAMP), 12 recombinase polymerase amplification (RPA) 13 and recombinase aided amplification (RAA), 14 due to their high sensitivity, simple operation and low cost. Of these methods, RAA has received an increasing attention recently since its ability to perform exponential amplification within 20 min at a lower reaction temperature (37–42 °C) by introducing recombinase and single-strand binding protein.…”

Section: Introductionmentioning

confidence: 99%

An automatic centrifugal system for rapid detection of bacteria based on immunomagnetic separation and recombinase aided amplification

Wang

et al. 2022

Lab Chip

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A Simplified Amplification‐Free Strategy with Lyophilized CRISPR‐CcrRNA System for Drug‐Resistant Salmonella Detection

Bao

Sun

et al. 2023

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With the widespread abuse of antibiotics, the problem of bacterial resistance is becoming increasingly serious, resulting in more than 700 000 deaths annually, worldwide. [1] Salmonella, one of the most dangerous and infectious foodborne pathogens, ranks second among pathogens that endanger public food safety. [2] Salmonella accounts for 41% of global diarrhea-related deaths. [1,3] The emergence of drug-resistant Salmonella has increased the danger to public health. Early diagnosis and prompt detection of drug resistance require a rapid, sensitive, and cost-effective detection program from the perspective of prevention, screening for drug-resistant bacteria, and management of food-related accidents. [4,5] Traditional culture-based antibiotic sensitivity testing (AST) methods are complicated and often require 2 to 7 days. [6] In contrast, nucleic acid detection of drug-resistance genes has significant advantages in terms of sensitivity and specificity. [7] The gold standard for nucleic acid detection, quantitative polymerase chain reaction (qPCR), can detect 10 CFU mL −1 of resistant bacteria within a few hours. [8,9] However, it still relies heavily on specialized equipment and expertise and is difficult to use outside a laboratory setting. [4] Lateral flow assay (LFA) is the most promising point-of-care testing (POCT) platform owing to its minimal device dependence, speed, sensitivity, and user-friendliness. [10,11] A visual readout LFA based on antigen capture with a faster and more accurate reading within 15 min was developed. [12] Unfortunately, this immunology-based pathogenic bacterial detection via LFA generally has low sensitivity and cannot be used for AST. [10,13] Combining the CRISPR/Cas system with LFA allows for sensitive and specific detection of targets while maintaining its fast and user-friendly features. For instance. Gootenberg et al [14] proposed a method called SHERLOCK for SARS-CoV-2 detection, [15] Joung et al. [13] demonstrated the direct detection of SARS-CoV-2 samples and further developed the SHINE. [16] All of the LFA-CRISPR methods mentioned above enable Drug resistance in pathogenic bacteria has become a major threat to global health. The misuse of antibiotics has increased the number of resistant bacteria in the absence of rapid, accurate, and cost-effective diagnostic tools. Here, an amplification-free CRISPR-Cas12a time-resolved fluorescence immunochromatographic assay (AFC-TRFIA) is used to detect drugresistant Salmonella. Multi-locus targeting in combination crRNA (CcrRNA) is 27-fold more sensitive than a standalone crRNA system. The lyophilized CRISPR system further simplifies the operation and enables one-pot detection. Induction of nucleic acid fixation via differentially charged interactions reduced the time and cost required for flowmetric chromatography with enhanced stability. The induction of nucleic acid fixation via differentially charged interactions reduces the time and cost required for flowmetric chromatography with enhanced stability. The platform developed for th...

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Loop-mediated isothermal amplification (LAMP) for rapid detection of Salmonella in foods based on new molecular targets

Cited by 31 publications

References 37 publications

Real-Time PCR Method for the Rapid Detection and Quantification of Pathogenic Staphylococcus Species Based on Novel Molecular Target Genes

Real-Time PCR Method for the Rapid Detection and Quantification of Pathogenic Staphylococcus Species Based on Novel Molecular Target Genes

An automatic centrifugal system for rapid detection of bacteria based on immunomagnetic separation and recombinase aided amplification

A Simplified Amplification‐Free Strategy with Lyophilized CRISPR‐CcrRNA System for Drug‐Resistant Salmonella Detection

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