Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F62157124%3A16270%2F17%3A43875823" target="_blank" >RIV/62157124:16270/17:43875823 - isvavai.cz</a>

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.jviromet.2017.02.017" target="_blank" >http://dx.doi.org/10.1016/j.jviromet.2017.02.017</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.jviromet.2017.02.017" target="_blank" >10.1016/j.jviromet.2017.02.017</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Popis výsledku v původním jazyce

Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TagMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97.1%, with consistent detection of 101 virus copies/mu l of nucleic acid extract. The low values of standard deviation and coefficient of variation indicate a high degree of reproducibility and repeatability. The diagnostic applicability of the assay was proven on 36 BFDV positive and 107 negative specimens of psittacine origin representing 28 species. The assay showed a 100% ability to detect distinct genetic variants of the virus. Our data suggest that the presented TaqMan real-time PCR represents a specific, sensitive and reliable assay facilitating the molecular detection of BFDV.

Název v anglickém jazyce

Development and evaluation of TaqMan real-time PCR assay for detection of beak and feather disease virus

Popis výsledku anglicky

Psittacine beak and feather disease (PBFD) is one of the most significant viral diseases in psittacine birds. The aim of the presented study was to develop a highly specific and sensitive TagMan real-time PCR assay for universal detection of beak and feather disease virus (BFDV). Primers and a hydrolysis probe were selected on the highly conserved regions belonging to the ORF1 of the BFDV genome which were identified by aligning 814 genomic sequences downloaded from the GenBank database. The evaluation of the reaction parameters suggested a reaction efficiency of 97.1%, with consistent detection of 101 virus copies/mu l of nucleic acid extract. The low values of standard deviation and coefficient of variation indicate a high degree of reproducibility and repeatability. The diagnostic applicability of the assay was proven on 36 BFDV positive and 107 negative specimens of psittacine origin representing 28 species. The assay showed a 100% ability to detect distinct genetic variants of the virus. Our data suggest that the presented TaqMan real-time PCR represents a specific, sensitive and reliable assay facilitating the molecular detection of BFDV.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

40301 - Veterinary science

Projekt

—

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Journal of Virological Methods

ISSN

0166-0934

e-ISSN

—

Svazek periodika

244

Číslo periodika v rámci svazku

JUN 2017

Stát vydavatele periodika

NL - Nizozemsko

Počet stran výsledku

6

Strana od-do

55-60

Kód UT WoS článku

000401677200010

EID výsledku v databázi Scopus

—