Combination of reverse transcription real-time polymerase chain reaction and antigen capture enzyme-linked immunosorbent assay for the detection of animals persistently infected with Bovine viral diarrhea virus.
J Vet Diagn Invest
; 23(1): 16-25, 2011 Jan.
Article
em En
| MEDLINE
| ID: mdl-21217023
ABSTRACT
Bovine viral diarrhea virus (BVDV) is an economically important pathogen of cattle. A successful control program requires early detection and removal of persistently infected (PI) animals. The objective of the current study was to develop, validate, and apply a cost-effective testing scheme for the detection of BVDV PI animals in exposed herds. Pooled samples were screened by using a real-time reverse transcription polymerase chain reaction (real-time RT-PCR), and individual positives were identified with an antigen capture enzyme-linked immunosorbent assay (ACE). The detection limits of the optimized real-time RT-PCR were 10 and 100 RNA copies per reaction for BVDV-1 and BVDV-2, respectively. The semiquantitative results of real-time RT-PCR and ACE or real-time RT-PCR and immunohistochemistry were moderately correlated. The threshold cycle of real-time RT-PCR performed on pooled samples was significantly correlated with the pool size (R(2) â=â 0.993). The least-cost pool sizes were 50 at a prevalence of 0.25-0.5% and 25 at a prevalence of 0.75-2.0%. By using the combined real-time RT-PCR and ACE procedure, 111 of 27,932 samples (0.4%) tested positive for BVDV. At this prevalence, cost reduction associated with the application of real-time RT-PCR and ACE ranged from 61% to 94%, compared with testing individual samples by ACE, immunohistochemistry, or real-time RT-PCR. Real-time RT-PCR screening also indicated that 92.94% of PI animals were infected with BVDV-1, 3.53% with BVDV-2, and 3.53% with both BVDV-1 and BVDV-2. Analysis of the 5'-untranslated region of 22 isolates revealed the predominance of BVDV-1b followed by BVDV-2a.
Buscar no Google
Base de dados:
MEDLINE
Assunto principal:
Doença das Mucosas por Vírus da Diarreia Viral Bovina
/
Ensaio de Imunoadsorção Enzimática
/
Portador Sadio
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Vírus da Diarreia Viral Bovina Tipo 1
/
Vírus da Diarreia Viral Bovina Tipo 2
Tipo de estudo:
Diagnostic_studies
/
Prevalence_studies
/
Risk_factors_studies
/
Screening_studies
Limite:
Animals
Idioma:
En
Ano de publicação:
2011
Tipo de documento:
Article