Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry

Bacterial leaf scorch, caused by Xylella fastidiosa, is a major threat to blueberry production in the southeastern United States. Management of this devastating disease is challenging and often requires early detection of the pathogen to reduce major loss. There are several different molecular and s...

Descripción completa

Detalles Bibliográficos
Autores principales: Waliullah, Sumyya, Hudson, Owen, Oliver, Jonathan E., Brannen, Phillip M., Ji, Pingsheng, Ali, Md Emran
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719857/
https://www.ncbi.nlm.nih.gov/pubmed/31479482
http://dx.doi.org/10.1371/journal.pone.0221903
_version_ 1783447995903115264
author Waliullah, Sumyya
Hudson, Owen
Oliver, Jonathan E.
Brannen, Phillip M.
Ji, Pingsheng
Ali, Md Emran
author_facet Waliullah, Sumyya
Hudson, Owen
Oliver, Jonathan E.
Brannen, Phillip M.
Ji, Pingsheng
Ali, Md Emran
author_sort Waliullah, Sumyya
collection PubMed
description Bacterial leaf scorch, caused by Xylella fastidiosa, is a major threat to blueberry production in the southeastern United States. Management of this devastating disease is challenging and often requires early detection of the pathogen to reduce major loss. There are several different molecular and serological detection methods available to identify the pathogen. Knowing the efficiency and suitability of these detection techniques for application in both field and laboratory conditions is important when selecting the appropriate detection tool. Here, we compared the efficiency and the functionality of four different molecular detection techniques (PCR, real-time PCR, LAMP and AmplifyRP® Acceler8™) and one serological detection technique (DAS-ELISA). The most sensitive method was found to be real-time PCR with the detection limit of 25 fg of DNA molecules per reaction (≈9 genome copies), followed by LAMP at 250 fg per reaction (≈90 copies), AmplifyRP® Acceler8™ at 1 pg per reaction (≈350 copies), conventional PCR with nearly 1.25 pg per reaction (≈ 440 copies) and DAS-ELISA with 1x10(5) cfu/mL of Xylella fastidiosa. Validation between assays with 10 experimental samples gave consistent results beyond the variation of the detection limit. Considering robustness, portability, and cost, LAMP and AmplifyRP® Acceler8™ were not only the fastest methods but also portable to the field and didn’t require any skilled labor to carry out. Among those two, AmplifyRP® Acceler8™ was faster but more expensive and less sensitive than LAMP. On the other hand, real-time PCR was the most sensitive assay and required comparatively lesser time than C-PCR and DAS-ELISA, which were the least sensitive assays in this study, but all three assays are not portable and needed skilled labor to proceed. These findings should enable growers, agents, and diagnosticians to make informed decisions regarding the selection of an appropriate diagnostic tool for X. fastidiosa on blueberry.
format Online
Article
Text
id pubmed-6719857
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-67198572019-09-16 Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry Waliullah, Sumyya Hudson, Owen Oliver, Jonathan E. Brannen, Phillip M. Ji, Pingsheng Ali, Md Emran PLoS One Research Article Bacterial leaf scorch, caused by Xylella fastidiosa, is a major threat to blueberry production in the southeastern United States. Management of this devastating disease is challenging and often requires early detection of the pathogen to reduce major loss. There are several different molecular and serological detection methods available to identify the pathogen. Knowing the efficiency and suitability of these detection techniques for application in both field and laboratory conditions is important when selecting the appropriate detection tool. Here, we compared the efficiency and the functionality of four different molecular detection techniques (PCR, real-time PCR, LAMP and AmplifyRP® Acceler8™) and one serological detection technique (DAS-ELISA). The most sensitive method was found to be real-time PCR with the detection limit of 25 fg of DNA molecules per reaction (≈9 genome copies), followed by LAMP at 250 fg per reaction (≈90 copies), AmplifyRP® Acceler8™ at 1 pg per reaction (≈350 copies), conventional PCR with nearly 1.25 pg per reaction (≈ 440 copies) and DAS-ELISA with 1x10(5) cfu/mL of Xylella fastidiosa. Validation between assays with 10 experimental samples gave consistent results beyond the variation of the detection limit. Considering robustness, portability, and cost, LAMP and AmplifyRP® Acceler8™ were not only the fastest methods but also portable to the field and didn’t require any skilled labor to carry out. Among those two, AmplifyRP® Acceler8™ was faster but more expensive and less sensitive than LAMP. On the other hand, real-time PCR was the most sensitive assay and required comparatively lesser time than C-PCR and DAS-ELISA, which were the least sensitive assays in this study, but all three assays are not portable and needed skilled labor to proceed. These findings should enable growers, agents, and diagnosticians to make informed decisions regarding the selection of an appropriate diagnostic tool for X. fastidiosa on blueberry. Public Library of Science 2019-09-03 /pmc/articles/PMC6719857/ /pubmed/31479482 http://dx.doi.org/10.1371/journal.pone.0221903 Text en © 2019 Waliullah et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Waliullah, Sumyya
Hudson, Owen
Oliver, Jonathan E.
Brannen, Phillip M.
Ji, Pingsheng
Ali, Md Emran
Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title_full Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title_fullStr Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title_full_unstemmed Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title_short Comparative analysis of different molecular and serological methods for detection of Xylella fastidiosa in blueberry
title_sort comparative analysis of different molecular and serological methods for detection of xylella fastidiosa in blueberry
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6719857/
https://www.ncbi.nlm.nih.gov/pubmed/31479482
http://dx.doi.org/10.1371/journal.pone.0221903
work_keys_str_mv AT waliullahsumyya comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry
AT hudsonowen comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry
AT oliverjonathane comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry
AT brannenphillipm comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry
AT jipingsheng comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry
AT alimdemran comparativeanalysisofdifferentmolecularandserologicalmethodsfordetectionofxylellafastidiosainblueberry