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A two-dimensional pooling approach towards efficient detection of parasitoid and pathogen DNA at low infestation rates.

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Title A two-dimensional pooling approach towards efficient detection of parasitoid and pathogen DNA at low infestation rates.
 
Creator Sint, D.
Sporleder, M.
Wallinger, C.
Zegarra, O.
Oehm, J.
Dangi, N.
Giri, Y.P.
Kroschel, J.
Traugott, M.
 
Subject MOLECULAR GENETICS
PATHOGENS
PARASITOIDS
 
Description Molecular methods are facing increasing application on a broad remit in ecological studies, including work on parasites, parasitoids and pathogens. These studies can entail the screening of large sample numbers to monitor the establishment of parasitoid and pathogen infection rates over extended periods of time. However, when infection or parasitism rates are low, testing many samples individually quickly becomes tedious and costly. To optimize the application of molecular diagnostics in such situations, we developed a pooling approach that is applicable to any host–pathogen and host–parasite/parasitoid system.
In a first step, host individuals are pooled along the two axes of a grid and screened for pathogen/parasite DNA via multiplex PCR. Already from these results a rough estimation of the pathogen/parasitism rate in the pooled samples is possible. To determine the exact infection/parasitism rate in the sample or if it is relevant to identify the respective infected/parasitized individuals, only those samples belonging to two pathogen/parasitoid DNA-positive pools need to be retested individually. The efficacy of the pooling approach was statistically and empirically evaluated for different infestation levels using three host–parasitoid systems and a newly developed diagnostic multiplex PCR assay.
The established multiplex PCR assay allows to reliably identifying three agro-economically important lepidopteran pest species (Phthorimaea operculella, Symmetrischema tangolias and Tuta absoluta) and five relevant parasitoids (Copidosoma koehleri, Apanteles subandinus, Orgilus lepidus, Dolichogenidea gelechiidivoris and Diadegma mollipla). Compared to individual testing and even to standard one-dimensional pooling approaches, the new method reduces the number of PCRs substantially. At parasitism rates
Peer Review
 
Date 2017-03-28T12:57:03Z
2017-03-28T12:57:03Z
2016-08
 
Type Journal Article
 
Identifier Sint, D.; Sporleder, M.; Wallinger, C.; Zegarra, O.; Oehm, J.; Dangi, N.; Giri, Y.P.; Kroschel, J.; Traugott, M. 2016. A two-dimensional pooling approach towards efficient detection of parasitoid and pathogen DNA at low infestation rates. Methods in Ecology and Evolution. (USA). ISSN 2041-210X. 7(12):1548–1557.
2041-210X
https://hdl.handle.net/10568/80538
 
Language en
 
Format 1548–1557
 
Source Methods in Ecology and Evolution