Development of a novel and rapid polymerase spiral reaction (PSR) assay to detect Salmonella in pork and pork products

Momin, Kasanchi M.; Milton, Arockiasamy Arun Prince; Ghatak, Sandeep; Thomas, Shiny; Priya, G. Bhuvana; Das, Samir; Shakuntala, I.; Sanjukta, Rajkumari; Puro, K.

doi:10.1016/j.mcp.2020.101510

Cited by 30 publications

(27 citation statements)

References 35 publications

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“…We considered the qPCR or culture assay as “gold standard” for fungal identification ( Momin et al, 2020 ). Then, we used these techniques to estimate positive detection rates, sensitivities, specificities, and positive predictive and negative predictive values of ddPCR in suspected candidemia.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

Chen

Ying-guang²,

Zhang³

et al. 2021

Front. Microbiol.

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Numerous studies have shown that droplet digital PCR (ddPCR) is a promising tool for the diagnosis of pathogens, especially in samples with low concentrations of pathogenic DNA. An early diagnosis of candidemia is critical for the effective treatment of patients. In this study, we evaluated the sensitivity and specificity of ddPCR assay for Candida DNA detection both in vitro by mixing fungal cells with human blood and in vivo by analyzing blood samples from infected mice and patients with suspected candidemia. The results showed that ddPCR assay could detect a minimum of 4.5 DNA copies per reaction in blood samples. ddPCR showed higher sensitivity and specificity for Candida DNA detection than traditional culture and quantitative PCR (qPCR) methods and also exhibited significantly better positive and negative predictive values than the culture and qPCR methods that were commonly used in clinical practice. Hence, our study demonstrates that ddPCR assay is a promising method for the timely diagnosis of candidemia and could be useful for monitoring the treatment of candidemia.

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Section: Methodsmentioning

confidence: 99%

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

Chen

Ying-guang²,

Zhang³

et al. 2021

Front. Microbiol.

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“…Drawbacks of LAT are the shorter shelf life of beads and qualitative nature (Mangal et al, 2016). (Momin et al, 2020). To achieve selective detection of viable cells, certain dyes like ethidium monoazide (EMA) or propidium monoazide (PMA) have been used with PCR, which is termed as viability PCR (Pan and Breidt, 2007).…”

Section: Immunological Methodsmentioning

confidence: 99%

“…RT-qPCR is the preferred method of choice for screening foodborne viruses in food. Isothermal based detection techniques can be visually interpreted, they obviate the necessity of thermal cycler and gel electrophoresis, thus making them rapid and simple (Momin et al, 2020). Many commercial LAMP-based kits are available for detecting egg/poultry borne pathogens , Campylobacter, Listeria, etc (Mangal et al, 2016).…”

Section: Immunological Methodsmentioning

confidence: 99%

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An appraisal of various pathogen detection methods in eggs and poultry

Milton

Priya

Momin

et al. 2020

JFAS

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To ensure safety in egg and poultry products, timely detection of pathogenic microbes is of paramount importance. This review offers an appraisal of different routinely used and novel emerging pathogen detection methods in egg, poultry and their products. Timely detection of pathogens is decisive to curtail outbreak risks, reduce hospitalisation, and provide product assurance. It will also reduce the cost of holding food products in cold storage and reduces product recalls. Some crucial issues need to be taken care of in choosing or developing a foodborne pathogen detection method. They are requirement of costly or sophisticated equipment, portability, trained personnel, viable but non-culturable bacteria (may give false-negative results), dead microbes (may give false-positive results), stressed or sub-lethally damaged bacteria and slow-growing microbes (require enrichment). In this review, the focus has been given on culture-based methods, nucleic acid-based methods, immunological methods and biosensor based methods. Keywords: Egg; poultry; detection methods; product assurance; safety.

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“…Salmonella is an important foodborne pathogen responsible for frequent gastroenteritis, intestinal illnesses, foodborne outbreaks and deaths worldwide (Milton et al 2018;Bi et al 2020;Momin et al 2020). Globally, approximately 1Á3 billion gastroenteritis cases are attributed to Salmonella every year with varying clinical presentations ranging from mild enteric infection to systemic diseases (Waldman et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Development of novel visual detection methodology for Salmonella in meat using saltatory rolling circle amplification

et al. 2021

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Aim:The aim of this study was to develop a saltatory rolling circle amplification (SRCA) assay for rapid, simple and visual detection of Salmonella in meat. Methods and Results: Saltatory rolling circle amplification assay was established using simple PCR primers targeting the invA gene of Salmonella enterica. The specificity of the SRCA assay was determined using 28 Salmonella and 15 non-Salmonella strains. The analytical sensitivity of the developed SRCA, conventional and real-time PCR assays were 70 fg, 7 pg and 700 fg S. enterica DNA per tube, respectively. The limit of detection (LoD) of the SRCA assay was 40 CFU per gram of meat without enrichment and 4 CFU per gram after including 6 h brief enrichment step. The detection limits of 40 CFU per gram and 4 CFU per gram of meat were achieved within 165 min and 9 h, respectively (including DNA extraction). To assess the real-world relevance of the SRCA assay, it was used to screen Salmonella from the field pork samples (n = 82). The same samples were also tested with culture (ISO 6579: 2002) method, conventional and real-time PCR assays. Using the developed assay with 6-h enrichment step, it could give accurate results as that of the culture method. Conclusions: The results of this study showed that the SRCA assay is a rapid, simple, sophisticated equipment-free and user-friendly method for accurate detection of Salmonella in meat foods. To our information, this is the first study to deploy SRCA assay for screening foods for Salmonella. Significance and Impact of the Study: The developed SRCA assay is costeffective, easy-to-perform and equipment-free; therefore, it has the potential to replace other molecular detection methods for regular screening of Salmonella in foods in field laboratories.

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Development of a novel and rapid polymerase spiral reaction (PSR) assay to detect Salmonella in pork and pork products

Cited by 30 publications

References 35 publications

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

Evaluation of Droplet Digital PCR Assay for the Diagnosis of Candidemia in Blood Samples

An appraisal of various pathogen detection methods in eggs and poultry

Development of novel visual detection methodology for Salmonella in meat using saltatory rolling circle amplification

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