A real-time recombinase polymerase amplification assay for fast and accurate detection of Ditylenchus destructor. (February 2022)

• Record Type: Journal Article
• Authors: Gao, Bo; Ma, Juan; Li, Xiuhua; Wang, Rongyan; Chen, Shulong
• Is Part Of: Molecular and cellular probes. Volume 61(2022)
• Publication Date: 2022-02
• Subjects: Ditylenchus destructor -- Plant-parasitic nematodes -- Real-time -- Recombinase polymerase amplification -- Detection; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2021.101788
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: Ditylenchus destructor is a plant-parasitic nematode that seriously infests sweet potato crop in China. Thus, fast and accurate detection of D. destructor in soil and plant tissue samples is of great significance. In this study, a real-time recombinase polymerase amplification (RPA) assay was developed for the rapid and accurate detection of D . destructor in various samples. The RPA assay could be easily operated and detected as low as 1/500 individual J4 nematode DNA per reaction in 20 min at 39 °C with high specificity. The assay meets the requirements of rapid detection prior to port quarantine as well as on-site real-time detection and can be applied to detect the parasite in soil and plant samples. The modified gDNA extraction method for a single nematode established in this study significantly reduced the time of detection and improved the applicability of the real-time RPA assay for on-site detection in different environments. The real-time RPA assay to detect D. destructor will be useful for epidemiological investigations in the field as well as for quarantine processes in the sweet potato and potato trade. Highlights: A real-time RPA assay with high sensitivity and specificity was developed to detect Ditylenchus destructor. The RPA assay can detect as low as 1/500 individual J4 nematode gDNA in 20 min. The RPA assay can directly detect D. destructor from the DNA of soil and plant tissue.