From laboratory to point of entry: development and implementation of a loop‐mediated isothermal amplification (LAMP)‐based genetic identification system to prevent introduction of quarantine insect species

BACKGROUND Rapid genetic on‐site identification methods at points of entry, such as seaports and airports, have the potential to become important tools to prevent the introduction and spread of economically harmful pest species that are unintentionally transported by the global trade of plant commod...

Full description

Saved in:
Bibliographic Details
Published in:Pest management science 2018-06, Vol.74 (6), p.1504-1512
Main Authors: Blaser, Simon, Diem, Hanspeter, von Felten, Andreas, Gueuning, Morgan, Andreou, Michael, Boonham, Neil, Tomlinson, Jennifer, Müller, Pie, Utzinger, Jürg, Frey, Jürg E, Bühlmann, Andreas
Format: Article
Language:English
Subjects:
Airports
Commodities
Cytochrome
Cytochrome-c oxidase
Deoxyribonucleic acid
DNA
DNA barcoding
Evaluation
Fruit flies
Gene sequencing
Identification methods
Identification systems
Insects
International trade
Laboratories
loop‐mediated isothermal amplification
Mitochondria
Nucleotide sequence
Pest control
Pests
plant health inspections
point‐of‐entry diagnostics
Primers
Quarantine
quarantine organisms
Species
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:BACKGROUND Rapid genetic on‐site identification methods at points of entry, such as seaports and airports, have the potential to become important tools to prevent the introduction and spread of economically harmful pest species that are unintentionally transported by the global trade of plant commodities. This paper reports the development and evaluation of a loop‐mediated isothermal amplification (LAMP)‐based identification system to prevent introduction of the three most frequently encountered regulated quarantine insect species groups at Swiss borders, Bemisia tabaci, Thrips palmi and several regulated fruit flies of the genera Bactrocera and Zeugodacus. RESULTS The LAMP primers were designed to target a fragment of the mitochondrial cytochrome c oxidase subunit I gene and were generated based on publicly available DNA sequences. Laboratory evaluations analysing 282 insect specimens suspected to be quarantine organisms revealed an overall test efficiency of 99%. Additional on‐site evaluation at a point of entry using 37 specimens performed by plant health inspectors with minimal laboratory training resulted in an overall test efficiency of 95%. During both evaluation rounds, there were no false‐positives and the observed false‐negatives were attributable to human‐induced manipulation errors. To overcome the possibility of accidental introduction of pests as a result of rare false‐negative results, samples yielding negative results in the LAMP method were also subjected to DNA barcoding. CONCLUSION Our LAMP assays reliably differentiated between the tested regulated and non‐regulated insect species within
ISSN:1526-498X
1526-4998
DOI:10.1002/ps.4866