Developing and Testing a Real-Time Polymerase Chain Reaction to Identify and Quantify Bovine Respiratory Syncytial Viruses

Nefedchenko, A.V.; Глотов, А.Г.; Koteneva, S.V.; Glotova, T.I.

doi:10.3103/s0891416820030052

Cited by 6 publications

(3 citation statements)

References 22 publications

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“…In addition to serology, we analysed nasal/oropharyngeal swabs and tissue samples for the presence of BRSV-RNA using real-time RT-PCR and RT-PCR, as previously used by others [ 31 , 41 , 56 , 57 , 58 ]. In this study, rapid and specific detection of BRSV-RNA, probability-based real-time RT-PCR targeting the N gene of BRSV was used [ 39 , 56 , 58 , 59 ]; however, other genes, such as the F and G genes, have been targeted to detect BRSV-RNA in previous studies [ 60 , 61 ]. In the present study, 277 nasal/oropharyngeal swabs and 37 lung samples were analysed by real-time RT-PCR, with ten swabs (3.6%) and four lungs (10.8%) found to be positive for BRSV-RNA.…”

Section: Discussionmentioning

confidence: 99%

Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey

Aydin,

Yilmaz,

Turan

et al. 2024

Pathogens

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Bovine respiratory syncytial virus (BRSV) is one of the most important respiratory pathogens of cattle. In this study, frequency of infection, analysis of variants, and the immune status of vaccinated and non-vaccinated cattle were studied. Blood (n = 162) and nasal/oropharyngeal (n = 277) swabs were collected from 62 cattle herds in Turkey. Lung samples (n = 37) were also taken from dead animals and abattoirs. Antibodies to BRSV were detected in 76 (46%) out of 162 sera. The antibody levels in the vaccinated and non-vaccinated groups were statistically significant. Among 277 nasal/oropharyngeal swabs and 37 lungs, ten nasal/oropharyngeal and four lung samples were positive for BRSV-RNA. BRSV-G gene sequences of 5 out of 14 RT-PCR positive samples showed that all viruses clustered as Group-III in phylogenetic analysis with 88–100% homology. Similarity with previous Turkish BRSVs was 89–98%, and that with BRSVs detected in the USA and Czechia was 89.47–93.12%. BRSV continues to circulate in Turkish cattle, and vaccination seems beneficial in preventing BRSV. The diversity of the BRSVs found in this study needs be considered in vaccination strategies.

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Section: Discussionmentioning

confidence: 99%

Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey

Aydin,

Yilmaz,

Turan

et al. 2024

Pathogens

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“…Immunofluorescent antibody (IF) and enzyme-linked immunosorbent (ELISA) assays, which have 47% and 60% sensitivity for BRSV, respectively, are serological tests that are used to identify viral infection [5]. The gold standard for BRSV diagnosis at the moment, reverse transcription-polymerase chain reaction (RT-PCR) study, offers 99% sensitivity and specificity [8]. Some veterinary diagnostic laboratories are now unable to perform real-time PCR on a regular basis.…”

Section: Introductionmentioning

confidence: 99%

Comparative Evaluation between Nested RT-PCR andMono-Screen Antigen ELISA for Diagnosis of Bovine Respiratory Syncytial Virus in cattle

Hussain,

Al-Farwachi

2024

Egyptian Journal of Veterinary Sciences

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T HE MOST frequent cause of bovine respiratory illness in the world is respiratory syncytial virus (BRSV) of cattle. The objective of the present investigation was to detect the bovine respiratory syncytial virus (BRSV) utilizing nested RT-PCR and a mono-screen antigenenzyme-immunosorbent assay (Antigen-ELISA). It also searched for to evaluate the success of the diagnostic techniques. A total of four hundred and fifty nasal swabs from suspected respiratory infection cattle of local breeds aged 1-4 years were obtained. Out of the Four hundred and fifty nasals wabs, 168(37.3%) samples were positive to BRSV using RT-PCR technique (P< 0.05), while 139 (30.8%) samples were positive to BRSV using direct ELISA. However, 29 (6.4%) animals were negative results for the direct ELISA test but positive results for the RT-PCR technique. Considering antigen ELISA as the gold standard, the sensitivity and specificity of RT-PCR technique were 82.70% and 97% respectively. Based on Kappa value was (0.838), a perfect agreement between the two diagnostic methods. In conclusion , the One Tube RT-PCR System Script RT-PCR Premix Kit had the best sensitivity, specificity, and agreement when compared to the mono screen Antigen enzyme-linked immunosorbent assay BRSV /Sandwich double well (Antigen -ELISA). This implies that BRSV infection is detected using both assays. Antigen-ELISA is a sensitive, inexpensive test that may be used for routine diagnosis.

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“…Serological tests such as immunofluorescent antibody (IF) and enzyme-linked immunosorbent (ELISA) assays are also used to detect viral infection, exhibiting 47% and 60% sensitivity for BRSV, respectively. By comparison, analysis using reverse transcription-polymerase chain reaction (RT-PCR), which is currently considered the gold standard for BRSV diagnosis, has 99% sensitivity and specificity [1,3,11]. More recently, analytical techniques such as nuclear magnetic resonance (NMR) and gas chromatography based mass spectrometry (GC-MS) have been used in blood plasma, blood serum, nasal secretions, and exhaled breath condensate to identify biomarkers related to BRDC to create new diagnostic tools that facilitate appropriate treatment [12][13][14][15][16][17].…”

Section: Introductionmentioning

confidence: 99%

Bovine Respiratory Syncytial Virus (BRSV) Infection Detected in Exhaled Breath Condensate of Dairy Calves by Near-Infrared Aquaphotomics

et al. 2022

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Bovine respiratory syncytial virus (BRSV) is a major contributor to respiratory disease in cattle worldwide. Traditionally, BRSV infection is detected based on non-specific clinical signs, followed by reverse transcriptase-polymerase chain reaction (RT-PCR), the results of which can take days to obtain. Near-infrared aquaphotomics evaluation based on biochemical information from biofluids has the potential to support the rapid identification of BRSV infection in the field. This study evaluated NIR spectra (n = 240) of exhaled breath condensate (EBC) from dairy calves (n = 5) undergoing a controlled infection with BRSV. Changes in the organization of the aqueous phase of EBC during the baseline (pre-infection) and infected (post-infection and clinically abnormal) stages were found in the WAMACS (water matrix coordinates) C1, C5, C9, and C11, likely associated with volatile and non-volatile compounds in EBC. The discrimination of these chemical profiles by PCA-LDA models differentiated samples collected during the baseline and infected stages with an accuracy, sensitivity, and specificity >93% in both the calibration and validation. Thus, biochemical changes occurring during BRSV infection can be detected and evaluated with NIR-aquaphotomics in EBC. These findings form the foundation for developing an innovative, non-invasive, and in-field diagnostic tool to identify BRSV infection in cattle.

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Developing and Testing a Real-Time Polymerase Chain Reaction to Identify and Quantify Bovine Respiratory Syncytial Viruses

Cited by 6 publications

References 22 publications

Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey

Molecular Characterisation and Antibody Response to Bovine Respiratory Syncytial Virus in Vaccinated and Infected Cattle in Turkey

Comparative Evaluation between Nested RT-PCR andMono-Screen Antigen ELISA for Diagnosis of Bovine Respiratory Syncytial Virus in cattle

Bovine Respiratory Syncytial Virus (BRSV) Infection Detected in Exhaled Breath Condensate of Dairy Calves by Near-Infrared Aquaphotomics

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