Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples

Frączyk, Magdalena; Woźniakowski, Grzegorz; Kowalczyk, Andrzej; Niemczuk, Krzysztof; Pejsak, Z.

doi:10.1111/lam.12569

Cited by 43 publications

(33 citation statements)

References 17 publications

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“…The CPA assay has been applied to detect other important veterinary pathogens, such as African swine fever virus (ASFV) and fowl adenovirus (FAdV). These results indicate that the sensitivity of this approach was equal to that of RT-PCR [36,37,38]. In our present study, it could detect as little as 50 fg of genomic DNA from B. motasi per reaction, which was equal to approximately 50 μl of 0.000005% parasitic erythrocytes.…”

Section: Discussionsupporting

confidence: 53%

Rapid Detection of Babesia motasi Responsible for Human Babesiosis by Cross-priming Amplification Combined with a Vertical Flow

Wang

Gao

Zhang

et al. 2020

Preprint

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Background Babesia motasi is known as an etiological agent of human and ovine babesiosis. Diagnosis of babesiosis is traditionally performed by microscopy, examining Giemsa-stained thin peripheral blood smears. Rapid detection and accurate identification of species are desirable for clinical care and epidemiological studies. Methods An easy to operate molecular method, which requires less capital equipment and incorporates cross priming amplification combined with a vertical flow (CPA-VF) visualization strip for rapid detection and identification of B. motasi. Results The CPA-VF targets the 18S rRNA gene and has a detection limit of 50 fg per reaction; no cross reaction was observed with other piroplasms infective to sheep or Babesia infective to humans. CPA-VF and real-time (RT)-PCR had sensitivities of 95.2% (95% confidence interval [CI], 78.1-99.4%) and 90.5% (72-97.6%) and specificities of 95.8 (80.5-99.5%) and 97.9 (83.5-99.9%), respectively, versus microscopy and nested (n) PCR combined with gene sequencing. The clinical performance of the CPA-VF assay was evaluated with field blood samples from sheep (n = 240) in Jintai county, Gansu Province, and clinical specimens (n = 492) obtained from patients bitten by ticks. Conclusions Our results indicate that the CPA-VF is a rapid, accurate, nearly instrument-free molecular diagnostic approach for identification of B. motasi. Therefore, it could be an alternative technique for epidemiological investigations and diagnoses of ovine and/or human babesiosis caused by B. motasi, especially in resource-limited regions.

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Section: Discussionsupporting

confidence: 53%

Rapid Detection of Babesia motasi Responsible for Human Babesiosis by Cross-priming Amplification Combined with a Vertical Flow

Wang

Gao

Zhang

et al. 2020

Preprint

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“…We then focus on the isothermal ampli cation assays, as they could work with a constant temperature device and don't rely on complex specialized equipment. We found that many kinds of isothermal ampli cation methods, such as RPA assay [17], loop-mediated isothermal ampli cation assay (LAMP) [18], cross-priming ampli cation (CPA) [19], and polymerase cross-linking spiral reaction (PCLSR) [20], had been established and widely used for the diagnosis of infectious disease. As an alternative to PCR-based method, RPA assay is an e cient tool for the rapid diagnosis of infectious disease, especially in resource-limited settings.…”

Section: Discussionmentioning

confidence: 99%

Development of a reverse transcription recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of classical swine fever virus

Fan¹,

Zhang²,

Chen³

et al. 2020

Preprint

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Background: Classical swine fever (CSF), caused by the infection of Classical swine fever virus (CSFV), is a highly contagious disease of pigs and has caused significant economic losses in the pig industry. The rapid and effective detection of CSFV would contribute to the eradication program against CSF. Thus, this study aimed to develop a rapid and simple method for CSFV detection.Results: Here, a new method based on reverse transcription recombinase polymerase amplification (RT-RPA) coupled with lateral flow dipstick (LFD) was established for detecting CSFV. The RPA assay could be completed within 20 min at 37oC and the results of the RPA assay could be visualized by LFD assay with the naked-eye inspection. This RT-RPA-LFD assay could be used to detect CSFV specifically, with no cross-reaction with other pathogens. Its detection limit was 10 pg of CSFV cDNA. Importantly, the RT-RPA-LFD assay has a good performance on the CSFV detection for clinical samples.Conclusions: The established RT-RPA-LFD assay greatly reduced the need for professional staff and sophisticated instruments and made disease detection convenient and feasible. This method would be useful for the prevention and control of CSF, especially in resource-limited settings.

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“…Using different enzymes with DNA polymerase activity, nucleic acid amplification is performed under isothermal condition without a thermal cycler. Based on the principle of this technology, several types of isothermal amplification method has developed and used to aide in determining the bio-sample such as loop-mediated isothermal amplification (LAMP), rolling circle amplification (RCA), cross-priming amplification (CPA) or helicase dependent amplification (HDA), etc 13171819…”

Section: Discussionmentioning

confidence: 99%

The development of loop-mediated isothermal amplification (LAMP) assays for the rapid authentication of five forbidden vegetables in strict vegetarian diets

Lee

Lien

et al. 2017

Sci Rep

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Plant-based food ingredients such as garlic, Chinese leek, Chinese onion, green onion and onion are widely used in many cuisines around the world. However, these ingredients known as the “five forbidden vegetables” (FFVs) are not allowed in some vegetarian diets. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed for the detection of FFVs using five respective LAMP primer sets. The specific primers targeted the ITS1-5.8S-ITS2 nuclear ribosomal DNA sequence regions among the five vegetables. The results demonstrated that the identification of FFVs using the newly developed LAMP assay is more sensitive than the traditional PCR method. Using pepper, basil, parsley, chili and ginger as references, established LAMP primer sets showed high specificity for the identification of the FFV species. Moreover, when FFVs were mixed with other plant ingredients at different ratios (100:0, 50:50, 20:80, 10:90, 5:95, 2:98, and 1:99), no cross-reactivity was evident using LAMP. Finally, genomic DNAs extracted from boiled and steamed FFVs in processed foods were used as templates; the performance of the LAMP reaction was not influenced using validated LAMP primers. Not only can FFV ingredients be identified but commercial foods containing FFVs can also be authenticated. This LAMP method will be useful for the authentication of FFVs in practical food markets in the future.

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Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples

Cited by 43 publications

References 17 publications

Rapid Detection of Babesia motasi Responsible for Human Babesiosis by Cross-priming Amplification Combined with a Vertical Flow

Rapid Detection of Babesia motasi Responsible for Human Babesiosis by Cross-priming Amplification Combined with a Vertical Flow

Development of a reverse transcription recombinase polymerase amplification assay with lateral flow dipstick for rapid detection of classical swine fever virus

The development of loop-mediated isothermal amplification (LAMP) assays for the rapid authentication of five forbidden vegetables in strict vegetarian diets

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