Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku

Olive (Olea europaea L.) is a natural host of Olive latent virus 1 (OLV-1) that was also found in citrus trees, tulips and tomatoes. OLV-1 was detected by next-generation sequencing (NGS) also in strawberry plants, and therefore a new real-time PCR system has been developed for its routine detection...

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Autor principal:	Lucie Valentová
Formato:	article
Lenguaje:	CS EN
Publicado:	VÝZKUMNÝ A ŠLECHTITELSKÝ ÚSTAV OVOCNÁŘSKÝ HOLOVOUSY s.r.o. 2021
Materias:	molecular methods accredited laboratory plant viruses sensitivity specificity repeatability Plant culture SB1-1110 Botany QK1-989
Acceso en línea:	https://doaj.org/article/d309ad3fb9e441f49d39f32934f2261a
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spelling	oai:doaj.org-article:d309ad3fb9e441f49d39f32934f2261a2021-11-22T06:10:58ZVývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku2695-1347https://doaj.org/article/d309ad3fb9e441f49d39f32934f2261a2021-11-01T00:00:00Zhttps://vpovsuo.cz/wp-content/uploads/2021/11/1_Valentova_VPO27_2_21.pdfhttps://doaj.org/toc/2695-1347Olive (Olea europaea L.) is a natural host of Olive latent virus 1 (OLV-1) that was also found in citrus trees, tulips and tomatoes. OLV-1 was detected by next-generation sequencing (NGS) also in strawberry plants, and therefore a new real-time PCR system has been developed for its routine detection. The designed diagnostic kit was validated following the PM 7/98 (4) standard issued by European and Mediterranean Plant Protection Organization (EPPO) to allow its use in accredited laboratories. Using a synthetic standard, the following performance parameters of the kit were set during validation: specificity, sensitivity, repeatability, and reproducibility. Based on available OLV-1 sequences and our own sequencing results, primers and probe were designed to specifically detect only OLV-1 isolates (100% specificity). Sensitivity of the detection was assessed to be 50 copies per reaction; 500 copies per reaction was used to calculate other parameters. Repeatability and reproducibility were expressed as a variation coefficient, and were found to be 1.05 % and 1.08 %, respectively. Obtained parameters proved that the real-time PCR system for the detection of OLV-1 in strawberries was functional, reliable, and could be used in accredited laboratories for valid OLV-1 diagnostic.Lucie ValentováVÝZKUMNÝ A ŠLECHTITELSKÝ ÚSTAV OVOCNÁŘSKÝ HOLOVOUSY s.r.o.articlemolecular methodsaccredited laboratoryplant virusessensitivityspecificityrepeatabilityPlant cultureSB1-1110BotanyQK1-989CSENVědecké Práce Ovocnářské, Vol 27, Iss 2, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	CS EN
topic	molecular methods accredited laboratory plant viruses sensitivity specificity repeatability Plant culture SB1-1110 Botany QK1-989
spellingShingle	molecular methods accredited laboratory plant viruses sensitivity specificity repeatability Plant culture SB1-1110 Botany QK1-989 Lucie Valentová Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
description	Olive (Olea europaea L.) is a natural host of Olive latent virus 1 (OLV-1) that was also found in citrus trees, tulips and tomatoes. OLV-1 was detected by next-generation sequencing (NGS) also in strawberry plants, and therefore a new real-time PCR system has been developed for its routine detection. The designed diagnostic kit was validated following the PM 7/98 (4) standard issued by European and Mediterranean Plant Protection Organization (EPPO) to allow its use in accredited laboratories. Using a synthetic standard, the following performance parameters of the kit were set during validation: specificity, sensitivity, repeatability, and reproducibility. Based on available OLV-1 sequences and our own sequencing results, primers and probe were designed to specifically detect only OLV-1 isolates (100% specificity). Sensitivity of the detection was assessed to be 50 copies per reaction; 500 copies per reaction was used to calculate other parameters. Repeatability and reproducibility were expressed as a variation coefficient, and were found to be 1.05 % and 1.08 %, respectively. Obtained parameters proved that the real-time PCR system for the detection of OLV-1 in strawberries was functional, reliable, and could be used in accredited laboratories for valid OLV-1 diagnostic.
format	article
author	Lucie Valentová
author_facet	Lucie Valentová
author_sort	Lucie Valentová
title	Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
title_short	Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
title_full	Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
title_fullStr	Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
title_full_unstemmed	Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku
title_sort	vývoj a validace diagnostického real-time pcr systému pro detekci olive latent viru 1 (olv-1) v rostlinách jahodníku
publisher	VÝZKUMNÝ A ŠLECHTITELSKÝ ÚSTAV OVOCNÁŘSKÝ HOLOVOUSY s.r.o.
publishDate	2021
url	https://doaj.org/article/d309ad3fb9e441f49d39f32934f2261a
work_keys_str_mv	AT lucievalentova vyvojavalidacediagnostickehorealtimepcrsystemuprodetekciolivelatentviru1olv1vrostlinachjahodniku
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Vývoj a validace diagnostického real-time PCR systému pro detekci Olive latent viru 1 (OLV-1) v rostlinách jahodníku

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