Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat
C. perfringens is a widespread foodborne pathogen and one of the major concerns in the meat industry. There is a need for a simple, rapid and equipment free detection system for C. perfringens as conventional anaerobic culture method is labour and resource intensive. Here, we applied a novel polymer...
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doaj-8f37ad9b445c45bbb7bac6d62e223daf2021-02-05T16:13:43ZengElsevierHeliyon2405-84402021-01-0171e05941Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meatA. Arun Prince Milton0Kasanchi M. Momin1Sandeep Ghatak2G. Bhuvana Priya3M. Angappan4Samir Das5K. Puro6R.K. Sanjukta7I. Shakuntala8A. Sen9B.K. Kandpal10Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, India; Corresponding author.Division of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, India; College of Agriculture, Central Agricultural University (Imphal), Kyrdemkulai, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaDivision of Animal Health, ICAR Research Complex for NEH Region, Umiam, Meghalaya, IndiaC. perfringens is a widespread foodborne pathogen and one of the major concerns in the meat industry. There is a need for a simple, rapid and equipment free detection system for C. perfringens as conventional anaerobic culture method is labour and resource intensive. Here, we applied a novel polymerase spiral reaction phenomenon to develop and evaluate an assay for effortless and visual detection of C. perfringens in meat foods employing pork as a representative model. Specificity of the assay was determined using 51 C perfringens and 20 non- C. perfringens strains. Analytical sensitivity of the developed test was 80 fg DNA per tube indicating 100 times more sensitivity than end-point PCR assay. The detection limits were 980 CFU/g and 9.8 × 104 CFU/g of pork for PSR and PCR assays, respectively. The operation time of the PSR assay including DNA extraction was 120 min. The developed PSR assay was accurate and effective in comparison to culture method, in detecting C. perfringens in 38 of 74 pork samples. Therefore the specificity, sensitivity, negative predictive value, positive predictive value and accuracy rate of the developed PSR assay were 100%. The developed PSR assay is easy to perform, rapid, affordable, permitting sophisticated-equipment free amplification and naked eye interpretation. This is the initial report in which the PSR assay was optimized for the detection of C. perfringens.http://www.sciencedirect.com/science/article/pii/S2405844021000463C. perfringensIsothermal amplificationPSRRapid detectionMeatEquipment-free |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
A. Arun Prince Milton Kasanchi M. Momin Sandeep Ghatak G. Bhuvana Priya M. Angappan Samir Das K. Puro R.K. Sanjukta I. Shakuntala A. Sen B.K. Kandpal |
spellingShingle |
A. Arun Prince Milton Kasanchi M. Momin Sandeep Ghatak G. Bhuvana Priya M. Angappan Samir Das K. Puro R.K. Sanjukta I. Shakuntala A. Sen B.K. Kandpal Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat Heliyon C. perfringens Isothermal amplification PSR Rapid detection Meat Equipment-free |
author_facet |
A. Arun Prince Milton Kasanchi M. Momin Sandeep Ghatak G. Bhuvana Priya M. Angappan Samir Das K. Puro R.K. Sanjukta I. Shakuntala A. Sen B.K. Kandpal |
author_sort |
A. Arun Prince Milton |
title |
Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat |
title_short |
Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat |
title_full |
Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat |
title_fullStr |
Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat |
title_full_unstemmed |
Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat |
title_sort |
development of a novel polymerase spiral reaction (psr) assay for rapid and visual detection of clostridium perfringens in meat |
publisher |
Elsevier |
series |
Heliyon |
issn |
2405-8440 |
publishDate |
2021-01-01 |
description |
C. perfringens is a widespread foodborne pathogen and one of the major concerns in the meat industry. There is a need for a simple, rapid and equipment free detection system for C. perfringens as conventional anaerobic culture method is labour and resource intensive. Here, we applied a novel polymerase spiral reaction phenomenon to develop and evaluate an assay for effortless and visual detection of C. perfringens in meat foods employing pork as a representative model. Specificity of the assay was determined using 51 C perfringens and 20 non- C. perfringens strains. Analytical sensitivity of the developed test was 80 fg DNA per tube indicating 100 times more sensitivity than end-point PCR assay. The detection limits were 980 CFU/g and 9.8 × 104 CFU/g of pork for PSR and PCR assays, respectively. The operation time of the PSR assay including DNA extraction was 120 min. The developed PSR assay was accurate and effective in comparison to culture method, in detecting C. perfringens in 38 of 74 pork samples. Therefore the specificity, sensitivity, negative predictive value, positive predictive value and accuracy rate of the developed PSR assay were 100%. The developed PSR assay is easy to perform, rapid, affordable, permitting sophisticated-equipment free amplification and naked eye interpretation. This is the initial report in which the PSR assay was optimized for the detection of C. perfringens. |
topic |
C. perfringens Isothermal amplification PSR Rapid detection Meat Equipment-free |
url |
http://www.sciencedirect.com/science/article/pii/S2405844021000463 |
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