Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat

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

doi:10.1016/j.heliyon.2021.e05941

Cited by 9 publications

(8 citation statements)

References 35 publications

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“…The high specificity reported here was probably because of highly specific primers for the A. pleuropneumoniae apxIVA gene (Zhou et al, 2008). Similar levels of sensitivity of real-time PCR and PSR have also been reported by other workers (Ashmi et al, 2021;Das et al, 2018;Gupta et al, 2017;Malla et al, 2018;Milton et al, 2021;Xu et al, 2020). And the technology is ten times more sensitive compared to traditional PCR (Milton et al, 2021).…”

Section: Resultssupporting

confidence: 85%

“…PSR was not only sensitive but also rapid, already giving results after 30 min of incubation. In most reports, the minimum detection time by PSR was 45–60 min (Ashmi et al ., 2021; Das et al ., 2018; Dutta et al ., 2019; Gupta et al ., 2017; Malla et al ., 2018; Milton et al ., 2021; Xu et al ., 2020).…”

Section: Resultsmentioning

confidence: 99%

“…The high specificity reported here was probably because of highly specific primers for the A. pleuropneumoniae apxIVA gene (Zhou et al ., 2008). Similar levels of sensitivity of real‐time PCR and PSR have also been reported by other workers (Ashmi et al ., 2021; Das et al ., 2018; Gupta et al ., 2017; Malla et al ., 2018; Milton et al ., 2021; Xu et al ., 2020). And the technology is ten times more sensitive compared to traditional PCR (Milton et al ., 2021).…”

Section: Resultsmentioning

confidence: 99%

“…The polymerase spiral reaction (PSR) is the most recent inclusion in the list of INAT, which can amplify the target DNA with one pair of primers and a single enzyme (Dong et al ., 2015; Liu et al ., 2015) and it is being adopted for detection of many microbial organisms due to its simplicity, specificity, sensitivity and user‐friendly application (Dutta et al ., 2019). So far, PSR tests have been utilized to identify a variety of animal pathogens all around the world (Ashmi et al ., 2021; Das et al ., 2018; Dutta et al ., 2019; Milton et al ., 2021; Xu et al ., 2020). To date, to the best of our knowledge, no approach has been made to develop a PSR technique for early detection of A. pleuropneumoniae in pigs, which we attempted in the work presented here.…”

Section: Introductionmentioning

confidence: 99%

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Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction

Sarkar

Roychoudhury

Kumar

et al. 2022

Letters in Applied Microbiology

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Significance and Impact of the Study: Actinobacillus pleuropneumoniae is the primary aetiological agent of contagious porcine pleuropneumonia associated with serious economic impact on pig husbandry worldwide. Diagnosis of the disease by existing techniques including isolation and identification of bacteria followed by serotyping, serological techniques, conventional PCR, real-time PCR and LAMP assays are cumbersome, time-consuming, costly and not suitable for rapid field application. A novel isothermal polymerase spiral reaction (PSR) is developed for the specific detection of A. pleuropneumoniae in porcine nasal swabs. The technique is highly sensitive, specific, cost-effective and applicable to the field condition for rapid diagnosis of the disease. To the best of our knowledge, this is the first-ever report on the development of PSR for specific detection of A. pleuropneumoniae.

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Section: Resultssupporting

confidence: 85%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction

Sarkar

Roychoudhury

Kumar

et al. 2022

Letters in Applied Microbiology

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“…PSR assay for clinical diagnosis of viral and bacterial diseases and food pathogens are highly specific to their respective target nucleic acid sequences ( Liu et al, 2015 ; Gupta et al, 2017 ; Malla et al, 2018 ; Ji et al, 2019 ; Milton et al, 2020 , 2021 ; Sharma et al, 2020 , 2022 ; Tomar et al, 2020 ; Maiti et al, 2022 ). The PSR assay reported in the present study is specific to T. palmi mtCOIII region and does not cross-react to predominant thrips species.…”

Section: Discussionmentioning

confidence: 99%

Development of a Polymerase Spiral Reaction-Based Isothermal Assay for Rapid Identification of Thrips palmi

Jangra

Ghosh

Mukherjee

et al. 2022

Front. Mol. Biosci.

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Thrips cause considerable economic losses to a wide range of food, feed, and forest crops. They also transmit several plant viruses. Being cryptic, it is often difficult to distinguish thrips species in crops and large consignments by conventional methods. Melon thrips (Thrips palmi Karny, Thysanoptera: Thripidae) is an invasive insect pest of vegetables, legumes, and ornamentals besides being vector to several viruses. It poses a threat to domestic and international plant biosecurity and can invade and establish in new areas. Here, we report a polymerase spiral reaction (PSR)-based isothermal assay for rapid, sensitive, specific, low-cost, and on-site detection of T. palmi. To the best of our knowledge, this is the first application of PSR in the identification of any insect species. A primer pair designed based on 3′-polymorphism of mtCOIII region can specifically identify T. palmi without any cross-reactivity with predominant thrips species. The assay uses crude lysate of a single thrips saving time and reagents involved in nucleic acid extraction. The presence of T. palmi is visualized by the appearance of bright fluorescence under ultraviolet light or a change in reaction color thus avoiding gel electrophoresis steps. The entire process can be completed in 70 min on-site using only an ordinary water bath. The assay is sensitive to detecting as little as 50 attograms of T. palmi template. The assay was validated with known thrips specimens and found to be efficient in diagnosing T. palmi under natural conditions. The described method will be useful for non-expert personnel to detect an early infestation, accidental introduction to a new area, restrict the spread of diseases and formulate appropriate management strategies.

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Evaluation of reverse transcriptase-polymerase spiral reaction assay for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2

Prerana

Pai

Anupama

et al. 2023

Clinica Chimica Acta

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Development of a novel polymerase spiral reaction (PSR) assay for rapid and visual detection of Clostridium perfringens in meat

Cited by 9 publications

References 35 publications

Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction

Rapid detection of Actinobacillus pleuropneumoniae targeting the apxIVA gene for diagnosis of contagious porcine pleuropneumonia in pigs by polymerase spiral reaction

Development of a Polymerase Spiral Reaction-Based Isothermal Assay for Rapid Identification of Thrips palmi

Evaluation of reverse transcriptase-polymerase spiral reaction assay for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2

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