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Published November 16, 2017 | Version v1
Conference paper Open

Real-time LAMP rapid diagnostic method for X. fastidiosa in plant material and insect vectors

Description

X. fastidiosa is a gram-negative insect-vectored bacterium has been recently detected in Italian olive trees severely affected by Olive Quick Decline Syndrome (OQDS). ELISA tests and Polymerase Chain Reaction (PCR) assays were largely used in the monitoring campaign of this pathogen. The aim of this study is to evaluate the new real time Loop-mediated isothermal amplification (LAMP) system for the detection of X. fastidiosa in host plants and insect vectors. The new detection system is composed of a portable instrument (icgene mini) and a ready to use diagnostic kit denominated “Xylella Screen Glow” (Enbiotech s.r.l.- Italy). Specificity and sensitivity of Xylella Screen Glow kit was compared with PCR and real-time quantitative PCR assays. Three different DNA extraction protocols and typologies of infected materials were also tested. The real-time LAMP system showed high specificity and a sensitivity as compared to the real-time qPCR and PCR assays. These results showed that “Xylella Screen Glow” kit with icgene portable LAMP instrument is highly sensitive and suitable for X. fastidiosa detection directly in the field. Therefore, this system could be applied on site, with high sensitivity and readability to prevent the movement of infected materials from X. fastidiosa contaminated area to the laboratories located in free areas; other possible application could be in quarantine station to intercept any infected material before crossing border and protect from a country from such dangerous quarantine pathogen. 

Notes

This work has received funding from the European Union's Horizon 2020 research and innovation programme under grant agreement N. 727987 "Xylella fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy XF-ACTORS".

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Additional details

Funding

XF-ACTORS – Xylella Fastidiosa Active Containment Through a multidisciplinary-Oriented Research Strategy 727987
European Commission