Development of an improved real time PCR for the detection of bovine leukaemia provirus nucleic acid and its use in the clarification of nconclusive serological test results
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
258-264
Numer tomu
189
Identyfikator DOI
Liczba arkuszy
Autorzy
Pozostali autorzy
+ 2
Słowa kluczowe
en
Bovine leukaemia virus
Proviral DNA
Real time PCR
Streszczenia
Język
en
Treść
With the aim to erradicate Enzootic Bovine Leukosis from Poland, a more sensitive real-time polymerase chain reaction was required and developed to detect proviral Bovine leukaemia virus (BLV) DNA, the causative agent of Enzootic Bovine Leukosis (EBL). This new method proved more sensitive for our needs, than the current protocols available in the public domain. DNA was extracted from peripheral blood leukocytes of 51 cattle, which had given rise to doubtful serological test results by ELISA, and from mesenteric lymph nodes of six cattle that were slaughtered as EBL suspect cases. Additionally, fourteen DNA samples were obtained from animals with a strong BLV antibody response by ELISA. All real-time data were compared to results obtained from three different nested PCR methods. All 14 strongly positive ELISA samples were positive in all PCR tests. The real-time assay in comparison to the conventional PCR methods detected 7.8% (4/51) more specimens positive for BLV nucleic acid and showed a detection limit down to one copy. These observations represent the first report in the value of using a real-time method to help elucidate the disease status of animals when inconclusive ELISA results are obtained in the diagnostic laboratory. Thus, this method should be recommended for use in countries which have implemented an EBL-eradication programme, where a low level of BLV infection is evident.
Cechy publikacji
ORIGINAL_ARTICLE
Inne
System-identifier
552460
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