Evaluation of a single-tube fluorogenic RT-PCR assay for detection of bovine respiratory syncytial virus in clinical samples

Hakhverdyan, Mikhayil; Hägglund, Sara; Larsen, Lars Erik; Belák, Sándór

doi:10.1016/j.jviromet.2004.09.016

Cited by 20 publications

(17 citation statements)

References 18 publications

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“…Indeed, none of the 14 heterologous bovine viruses associated with bovine respiratory disease were detected, attesting to the high specificity of this assay. The detection limit obtained in the present study was 0.1 TCID 50 , which is comparable to the detection limit of previously reported real-time RT-PCR using the same primers and probe, 3 but 10 and 100 times more sensitive than real-time RT-PCR targeting the fusion gene 7 and conventional RT-PCR. 11 The reproducibility of the assay estimated on 10 runs was high with CVs lower than 5%.…”

supporting

confidence: 84%

Evaluation of a Commercial Real-Time Reverse Transcription Polymerase Chain Reaction Kit for the Diagnosis of Bovine Respiratory Syncytial Virus Infection

Timsit¹,

Maingourd

Dréan³

et al. 2010

J VET Diagn Invest

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Abstract. Recently a commercial real-time reverse transcription polymerase chain reaction (RT-PCR) kit has been marketed for the detection of Bovine respiratory syncytial virus (BRSV). However, diagnostic interpretation of the results of this kit requires its comparison to commonly used methods. Therefore, the objective of this study was to evaluate the performance of this kit in comparison with the conventional direct fluorescent antibody test (FAT). Twenty BRSV strains and 14 heterologous bovine viruses were used to check the kit's sensitivity and specificity. The efficiency and detection limit of the kit were determined by testing dilution series of a BRSV strain. In conclusion, the kit evaluated in this study was sensitive, specific, and had a low threshold of detection. Furthermore, the use of this kit instead of FAT allows an improvement of the sensitivity for the detection of BRSV in clinical samples.

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supporting

confidence: 84%

Evaluation of a Commercial Real-Time Reverse Transcription Polymerase Chain Reaction Kit for the Diagnosis of Bovine Respiratory Syncytial Virus Infection

Timsit¹,

Maingourd

Dréan³

et al. 2010

J VET Diagn Invest

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“…DNA and RNA were extracted using DNeasy Blood & Tissue Kit (Qiagen) and RNeasy Mini kit (Qiagen) according to the manufacturer's instructions. Viral real‐time PCR was performed on 34 lung samples targeting common viruses including BHV1, BRSV, BVDV, BPIV3 and BCV following standardized protocols (Decaro et al, ; Hakhverdyan, Hagglund, Larsen, & Belak, ; Horwood & Mahony, ; Mahlum et al, ; Wang et al, ). Real‐time PCR for M. bovis was performed on 120 lung samples (Clothier et al, ).…”

Section: Methodsmentioning

confidence: 99%

The pulmonary virome, bacteriological and histopathological findings in bovine respiratory disease from western Canada

Zhang

Hill

Godson

et al. 2019

Transbound Emerg Dis

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The aetiology and pathogenesis of bovine respiratory disease (BRD) are complex and involve the interplay of infectious agents, management and environmental factors. Previous studies of BRD focused on ante‐mortem samples from the upper respiratory tract and identified several unconventional viruses. The lung, however, is the primary location where significant BRD lesions are usually found and is a common post‐mortem diagnostic specimen. In this study, results of high‐throughput virome sequencing, bacterial culture, targeted real‐time PCR and histological examination of 130 bovine pneumonic lungs from western Canadian cattle were combined to explore associations of microorganisms with different types of pneumonia. Fibrinous bronchopneumonia (FBP) was the predominant type of pneumonia (46.2%, 60/130) and was associated with the detection of Mannheimia haemolytica. Detection of Histophilus somni and Pasteurella multocida was associated with suppurative bronchopneumonia (SBP) and concurrent bronchopneumonia and bronchointerstitial pneumonia (BP&BIP), respectively. Sixteen viruses were identified, of which bovine parvovirus 2 (BPV2) was the most prevalent (11.5%, 15/130) followed by ungulate tetraparvovirus 1 (UTPV1, 8.5%, 11/130) and bovine respiratory syncytial virus (BRSV, 8.5%, 11/130). None of these viruses, however, were significantly associated with a particular type of pneumonia. Unconventional viruses such as influenza D virus (IDV) and bovine rhinitis B virus (BRBV) were detected, although sparsely, consistent with our previous findings in upper respiratory tract samples. Taken together, our results show that while virus detection in post‐mortem lung samples is of relatively little diagnostic value, the strong associations of H. somni and M. haemolytica with SBP and FBP, respectively, indicate that histopathology can be useful in differentiating bacterial aetiologies.

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“…The samples were collected by the local veterinarians. This study included 30 samples from 19 dairy herds and 11 feedlot herds that were positive for BRSV by a real-time RT-PCR assay targeting the F gene [29] or by a gel-based PCR targeting the G gene [30]. During the study period there was a major epizootic of BRSV in the winter months of 2010/2011 in the county of Västerbotten, affecting at least 30 herds (area AC, Fig.…”

Section: Methodsmentioning

confidence: 99%

Phylogenetic analysis of bovine respiratory syncytial viruses from recent outbreaks in feedlot and dairy cattle herds

et al. 2011

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Bovine respiratory syncytial virus (BRSV) is one of the major causes of bovine respiratory disease worldwide. In order to study the molecular epidemiology of the virus, samples from 30 BRSV outbreaks in cattle herds located in different parts of Sweden were collected from 2007 to 2011. The samples were analyzed by PCR, and the glycoprotein (G) gene was sequenced. BRSV was detected in outbreaks of respiratory disease in both dairy and feedlot herds most often during the winter period but also during the summer months (May to August). This indicates that circulation of the virus between herds occurs throughout the year. Comparative sequence analysis revealed a high degree (more than 94.5%) of sequence identity among the collected strains. Phylogenetic analysis showed that 29 out of the 30 strains formed a unique clade. Identical sequences found in herds sampled within a few months' time suggested that these herds were part of a common transmission chain. One strain from a single outbreak in a herd in southern Sweden clustered with Danish strains and showed a distant relationship to the rest of the Swedish strains. Further studies are highly warranted to clarify the inter-herd transmission routes of BRSV. Such knowledge is essential for the control of the spread of this virus between herds, regions and even countries.

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Evaluation of a single-tube fluorogenic RT-PCR assay for detection of bovine respiratory syncytial virus in clinical samples

Cited by 20 publications

References 18 publications

Evaluation of a Commercial Real-Time Reverse Transcription Polymerase Chain Reaction Kit for the Diagnosis of Bovine Respiratory Syncytial Virus Infection

Evaluation of a Commercial Real-Time Reverse Transcription Polymerase Chain Reaction Kit for the Diagnosis of Bovine Respiratory Syncytial Virus Infection

The pulmonary virome, bacteriological and histopathological findings in bovine respiratory disease from western Canada

Phylogenetic analysis of bovine respiratory syncytial viruses from recent outbreaks in feedlot and dairy cattle herds

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