Rapid visualized isothermal nucleic acid testing of Vibrio parahaemolyticus by polymerase spiral reaction

He, Shiyu; Jang, Hongbo; Zhao, Chao; Xu, Kun; Wang, Juan; Pang, Bo; Si, Xiaoxue; Jin, Minghua; Song, Xiaohong; Li, Juan

doi:10.1007/s00216-019-02209-y

Cited by 27 publications

(10 citation statements)

References 24 publications

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“…KT327162.1), which has been confirmed by a series of reliable molecular biology references (Li W. et al, 2019;Richter et al, 2019). After performing blast sequence alignment in NCBI, two sets of primers were designed using the conserved sequence of the VP1 coding region as the target sequence (He et al, 2019), including primary primers (FT and BT) and accelerated primers (LF and LB). The primary primers were used for binding and folding with the target sequence, and the accelerated primers were used to accelerate the reaction process and achieve rapid detection.…”

Section: Design Of Plasmids and Primersmentioning

confidence: 96%

“…PSR combines the advantages of loop-mediated isothermal amplification (LAMP), without complicated heterothermic apparatus, and simple PCR primer design for rapid and effective detection of pathogens. Since its emergence, PSR has been applied to the detection of bacteria and double-stranded DNA viruses (Das et al, 2018;Malla et al, 2018;He et al, 2019). However, Coxsackievirus is a member of the Picornaviridae family of non-enveloped single-stranded RNA viruses, whose amplification involves a complex process of reverse transcription.…”

Section: Introductionmentioning

confidence: 99%

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A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

Huang

Yan

et al. 2020

Front. Microbiol.

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Hand, foot, and mouth disease (HFMD) is a common viral illness affecting infants and children that is usually caused by Coxsackievirus A16 (CVA-16). To diagnose HFMD, we developed a method for rapid detection of CVA-16 based on reverse transcriptionpolymerase spiral reaction (RT-PSR). We used two pairs of primers that specifically recognize the conserved sequences of VP1 coding region of CVA-16, and template RNA was reverse transcribed and amplified in a single tube under isothermal conditions, total reaction time could be reduced to less than 40 min. The detection limit of this method was between 2.4 × 10 2 and 2.4 × 10 1 copies/µl with excellent specificity. To test the clinical applicability of the method, 40 clinical stool samples were analyzed using RT-PSR and quantitative reverse transcription-polymerase chain reaction, and comparison showed that the coincidence rate was 100%. Compared with other similar detection methods, RT-PSR requires less time, simpler operation, and lower cost. These results prove that our novel, simple, and reliable isothermal nucleic acid testing assay has potential application for clinical detection of CVA-16.

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Section: Design Of Plasmids and Primersmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

Huang

Yan

et al. 2020

Front. Microbiol.

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“…PSR was developed for the detection of bacteria such as Salmonella and Vibrio parahaemolyticus. 36,37 For the detection of viruses, Gupta et al 38 developed PSR assay for the detection of canine parvovirus 2 and reported that the assay is more sensitive as compared to conventional PCR. RT-PSR assay was developed for the detection of RNA viruses like porcine epidemic diarrhoea virus (PEDV).…”

Section: Polymerase Spiral Reactionmentioning

confidence: 99%

Isothermal amplification‐based assays for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2: Opportunities and recent developments

Maiti

Anupama

Rai

et al. 2021

Reviews in Medical Virology

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Summary The coronavirus disease 2019 (COVID‐19) is a global pandemic caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). To date, the virus has been detected in 219 countries of the world. Therefore, managing the disease becomes the priority, in which detecting the presence of the virus is a crucial step. Presently, real‐time RT polymerase chain reaction (RT‐qPCR) is considered a gold standard nucleic acid amplification test (NAAT). The test protocol of RT‐qPCR is complicated, places high demands on equipment, testing reagents, research personnel skills and is expensive. Therefore, simpler point‐of‐care (POC) tests are needed to accelerate clinical decision‐making and take some of the workload from centralized test laboratories. Various isothermal amplification‐based assays have been developed for the sensitive detection of different microorganisms, and recently some of them have been applied for detection of SARS‐CoV‐2. These do not require any programable thermocycler, can produce the results in a single temperature, and therefore, are considered simple. Unlike RT‐qPCR, these methods are highly sensitive, specific, less time‐consuming, simple and affordable, and can be used as POC diagnostic kit for COVID‐19. In this review, we have discussed the potential of isothermal amplification‐based assays as an alternative to RT‐qPCR for the detection of SARS‐CoV‐2.

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“…Methods for detecting V. parahaemolyticus are mainly based on molecular biology and immunological techniques. Traditional molecular biology‐based detection methods include polymerase chain reaction (PCR) (He et al, 2020; Zeng et al, 2020) and real‐time PCR (He et al, 2018; Yoon et al, 2019), and more recent methods include loop‐mediated isothermal amplification (LAMP) (Anupama et al, 2019; Kampeera et al, 2019; Lee et al, 2020), rolling circle amplification (RCA) (Kampeera et al, 2019; Song et al, 2019) and recombinase polymerase amplification (RPA) (Geng et al, 2019; Jiang et al, 2020; Yang et al, 2018). Such methods have advantages in terms of detection time and sensitivity but have requirements for costly equipment and trained operators, which limit their application in low‐resource settings.…”

Section: Introductionmentioning

confidence: 99%

Detection of trh⁺Vibrio parahaemolyticus in seafood by lac dye coloration‐based label‐free lateral flow immunoassay strip

Liu

Tian

et al. 2022

Journal of Fish Diseases

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Vibrio parahaemolyticus (V. parahaemolyticus) is an important foodborne pathogen known to cause severe enteric disease. Thus, timely detection of V. parahaemolyticus in seafood is crucial to prevent food poisoning and reduce economic losses. Traditional lateral flow immunoassay strips (LFIS) required good labelling materials and pairing of two antibodies, which made them costly and difficult to manufacture. In this study, a label‐free and lac dye coloration‐based LFIS (LD‐LFIS) to detect trh+ V. parahaemolyticus was developed for the first time. Lac dye was used to stain V. parahaemolyticus, and LFIS was used to detect stained bacteria. Dimethyl sulphoxide (DMSO) and simultaneous mordanting were chosen as the best solvent and the best staining method for lac dye. In addition, three mordants [KAl(SO4)2·12H2O, NH4Fe(SO4)2·12H2O, and AlCl3·6H2O] were selected to improve dyeing efficiency. The detection limit of LD‐LFIS was 3.9 × 105 CFU/ml when NH4Fe(SO4)2·12H2O was used as mordant. Feasibility of the LD‐LFIS method was verified by detecting trh+ V. parahaemolyticus in true and spiked seafood samples. The method developed in this study is expected to reduce restrictions on labelling materials and pairing of two antibodies on LFIS, and proposes a novel idea for the rapid detection of V. parahaemolyticus in seafood.

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Rapid visualized isothermal nucleic acid testing of Vibrio parahaemolyticus by polymerase spiral reaction

Cited by 27 publications

References 24 publications

A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

Isothermal amplification‐based assays for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2: Opportunities and recent developments

Detection of trh⁺Vibrio parahaemolyticus in seafood by lac dye coloration‐based label‐free lateral flow immunoassay strip

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Rapid visualized isothermal nucleic acid testing of Vibrio parahaemolyticus by polymerase spiral reaction

Cited by 27 publications

References 24 publications

A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

A Reverse Transcription-Polymerase Spiral Reaction (RT-PSR)-Based Rapid Coxsackievirus A16 Detection Method and Its Application in the Clinical Diagnosis of Hand, Foot, and Mouth Disease

Isothermal amplification‐based assays for rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2: Opportunities and recent developments

Detection of trh+Vibrio parahaemolyticus in seafood by lac dye coloration‐based label‐free lateral flow immunoassay strip

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Detection of trh⁺Vibrio parahaemolyticus in seafood by lac dye coloration‐based label‐free lateral flow immunoassay strip