Development of a primer-probe energy transfer based real-time PCR for detection of influenza virus in pigs
PBN-AR
Instytucja
Państwowy Instytut Weterynaryjny - Państwowy Instytut Badawczy
Informacje podstawowe
Główny język publikacji
en
Czasopismo
JOURNAL OF VIROLOGICAL METHODS
ISSN
0166-0934
EISSN
Wydawca
ELSEVIER SCIENCE BV
DOI
URL
Rok publikacji
2013
Numer zeszytu
2
Strony od-do
228-233
Numer tomu
187
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Autorzy
Pozostali autorzy
+ 1
Słowa kluczowe
en
Primer–probe energy transfer
Swine influenza virus
PCR
Streszczenia
Język
en
Treść
Swine influenza virus (SIV) causes a contagious and requiring official notification disease of pigs and humans. In this study, a real-time reverse transcription-polymerase chain reaction (RT-PCR) assay based on primer–probe energy transfer (PriProET) for the detection of SIV RNA was developed. The assay uses matrix gene-specific primers and an Oregon Green-labeled fluorescent probe and was employed for the detection of SIV in clinical samples to identify outbreaks and to monitor the prevalence of disease. The PriProET technology was used to obtain a probe melting profile for confirmation of the specific product amplification. The assay is specific for influenza virus with a sensitivity of detection limit of approximately 10 copies of RNA by PCR. Based on serial dilutions of SIV, the detection limit of the assay was approximately 0.003 TCID50/ml for H1N1 A/Swine/Poland/KPR9/2004 virus. The PriProET RT-PCR was suitable for the detection of SIV RNA isolated directly from clinical samples. The assay detected SIV RNA in pre-clinical swab samples as early as 2 days post-infection (dpi). The PriProET RT-PCR assay is an alternative to the existing diagnostic assays and could have enhanced applicability for clinical diagnosis.
Cechy publikacji
ORIGINAL_ARTICLE
Inne
System-identifier
552160
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