Method for Detection of Extraneous Active Bovine Viral Diarrhoea Virus and Classical Swine Fever Virus in Animal Viral Vaccines by RT-PCR, Which Amplify Negative-strand Viral RNA in Infected Cells

Biologicals ◽  
2002 ◽  
Vol 30 (1) ◽  
pp. 27-35 ◽  
Author(s):  
Hiroshi Aoki ◽  
Tomoaki Shimazaki ◽  
Chikako Takahashi ◽  
Yoshimasa Sasaki ◽  
Shoko Suzuki ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Bovine Viral Diarrhoea Virus ◽  
Fever Virus ◽  
Viral Rna ◽  
Bovine Viral Diarrhoea ◽  
Rt Pcr ◽  
Negative Strand ◽  
Diarrhoea Virus ◽  
Infected Cells

Storage of bovine viral diarrhoea virus samples on filter paper and detection of viral RNA by a RT-PCR method

2001 ◽  
Vol 92 (1) ◽  
pp. 19-22 ◽  
Author(s):  
Štefan Vilček ◽  
Ladislav Strojny ◽  
Branislav Ďurkovič ◽  
Wigbert Rossmanith ◽  
David Paton
Keyword(s):  
Filter Paper ◽  
Bovine Viral Diarrhoea Virus ◽  
Viral Rna ◽  
Bovine Viral Diarrhoea ◽  
Rt Pcr ◽  
Diarrhoea Virus ◽  
Pcr Method

scholarly journals Evaluation of diagnostic methods to distinguish between calves persistently and transiently infected with bovine viral diarrhoea virus in respect to the presence of maternal antibodies

2013 ◽  
Vol 57 (3) ◽  
pp. 311-317
Author(s):  
Magdalena Larska ◽  
Aleksandra Kuta ◽  
Mirosław P. Polak
Keyword(s):  
Virus Detection ◽  
Bovine Viral Diarrhoea Virus ◽  
Diagnostic Methods ◽  
Viral Rna ◽  
Maternal Antibodies ◽  
Bovine Viral Diarrhoea ◽  
Rt Pcr ◽  
Serum Samples ◽  
Virus Genotype ◽  
Diarrhoea Virus

Abstract Two issues concerning virus detection and identification of persistently infected (PI) cattle were analysed in the study: 1) interference by maternal antibodies and 2) discrimination between PI and transiently infected (TI) animals. Antigen ELISA and RT-PCR based methods were compared using serum samples from natural and experimental PI and TI calves. RT-PCR and realtime RT-PCR using primers within 5’UTR region were more sensitive in detecting PI animals than Erns and NS3 antigen capture ELISAs, and they were not influenced by the presence of colostral antibodies in serum or by bovine viral diarrhoea virus genotype. The serum samples with Ct values ≤ 29.10 (corresponding to 104.87 viral RNA copies/μL) identified PI animals with 100% probability, while all samples with Ct values > 32.06 (corresponding to viral RNA load below 104 copies/μL) indicated TI status. The samples with Ct values between 29.10 and 32.06 (17.2% of PI and 11.5% of TI) should be considered as PI suspect and retested.

scholarly journals Short communication: Stability and integrity of classical swine fever virus RNA stored at room temperature

2017 ◽  
Vol 15 (3) ◽  
pp. e05SC03 ◽  
Author(s):  
Damarys Relova ◽  
Lester J. Pérez ◽  
Liliam Ríos ◽  
Liani Coronado ◽  
Yoandry Hinojosa ◽  
...  
Keyword(s):  
Classical Swine Fever Virus ◽  
Room Temperature ◽  
Storage Temperature ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Viral Rna ◽  
Nucleotide Sequence Analysis ◽  
Original Material ◽  
Rt Pcr ◽  
The Stability

Worldwide cooperation between laboratories working with classical swine fever virus (CSFV) requires exchange of virus isolates. For this purpose, shipment of CSFV RNA is a safe alternative to the exchange of infectious material. New techniques using desiccation have been developed to store RNA at room temperature and are reported as effective means of preserving RNA integrity. In this study, we evaluated the stability and integrity of dried CSFV RNA stored at room temperature. First, we determined the stability of CSFV RNA covering CSFV genome regions used typically for the detection of viral RNA in diagnostic samples by reverse transcription-polymerase chain reaction (RT-PCR). To this end, different concentrations of in vitro-transcribed RNAs of the 5’-untranslated region and of the NS5B gene were stored as dried RNA at 4, 20, and 37oC for two months. Aliquots were analyzed every week by CSFV-specific quantitative real-time RT-PCR. Neither the RNA concentration nor the storage temperature did affect CSFV RNA yields at any of the time evaluated until the end of the experiment. Furthermore, it was possible to recover infectious CSFV after transfection of SK-6 cells with dried viral RNA stored at room temperature for one week. The full-length E2 of CSFV was amplified from all the recovered viruses, and nucleotide sequence analysis revealed 100% identity with the corresponding sequence obtained from RNA of the original material. These results show that CSFV RNA stored as dried RNA at room temperature is stable, maintaining its integrity for downstream analyses and applications.

scholarly journals Monitoring of Bovine Viral Diarrhoea Virus (Bvdv) Infection in Polish Dairy Herds Using Bulk Tank Milk Samples

2013 ◽  
Vol 57 (2) ◽  
pp. 149-156 ◽  
Author(s):  
Aleksandra Kuta ◽  
Mirosław P. Polak ◽  
Magdalena Larska ◽  
Jan F. Żmudziński
Keyword(s):  
Bovine Viral Diarrhoea Virus ◽  
Viral Rna ◽  
Dairy Herds ◽  
Bovine Viral Diarrhoea ◽  
Rt Pcr ◽  
Bulk Tank Milk ◽  
Diarrhoea Virus ◽  
The Status ◽  
Bulk Tank ◽  
Antibody Levels

Abstract The aim of the study was to evaluate the status of bovine viral diarrhoea virus (BVDV) infection in selected dairy herds in Poland with the use of commercial enzyme linked immonosorbent assay for the detection of specific antibodies (BVDV-Ab ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) for the detection of viral RNA, using bulk tank milk (BTM) samples. Two hundred and thirty-one samples of BTM were collected from 99 dairy herds in Poland. The herds were divided into four different classes according to the Swedish system of classification. The results showed that 70.7% of herds were BVDV antibodypositive. High levels of antibodies in 52.85 % (37 herds in class 3) of all antibody positive herds indicated acute BVDV infection. Thirty five samples with the highest antibody levels were tested by RT-PCR and five of them were positive for viral RNA. Dairy herds in Poland have high levels of antibodies against BVDV in BTM. Since no vaccination was implemented in the herds tested, high seroprevalence of BVDV antibodies in cattle indicates the widespread of BVDV infection in Polish cattle.

A survey for bvdv antibodies in cattle farms in Slovakia and genetic typing of bvdv isolates from imported animals

2003 ◽  
Vol 51 (2) ◽  
pp. 229-236 ◽  
Author(s):  
Š. Vilček ◽  
Jana Mojžišová ◽  
Viera Bajová ◽  
Š. Paulík ◽  
L. Strojný ◽  
...  
Keyword(s):  
Bovine Viral Diarrhoea Virus ◽  
Computer Assisted ◽  
Serological Survey ◽  
Bovine Viral Diarrhoea ◽  
Rt Pcr ◽  
Serum Samples ◽  
Genetic Typing ◽  
Diarrhoea Virus ◽  
Pcr Products ◽  
Cattle Farms

A serological survey for bovine viral diarrhoea virus (BVDV) antibodies on a collection of 1295 serum samples obtained from 6-12 months old cattle originating from 45 farms in Slovakia was carried out. On 13 farms more than 90% of the examined animals were seropositive, on 14 farms 71-90% seroprevalence was observed, on 13 farms only 50-70% animals were found to be positive for BVDV antibodies, while the remaining 5 farms showed fewer than 50% seropositive animals. The average incidence of BVDV antibodies (around 70%) was similar as determined 30 years ago. Of 84 serum samples from seronegative animals originating from 14 farms in which 70-98% seropositivity was observed, six were positive in Ag-BVDV ELISA indicating persistently infected (PI) cattle. On a farm to which animals were imported from abroad, a BVD outbreak was observed. Of 110 animals tested, four were positive in Ag-ELISA indicating the presence of PI cattle on this farm. Genetic typing of two isolates from imported animals performed by RT-PCR (324/326 primers from 5´-UTR), sequencing of PCR products and computer-assisted phylogenetic analysis revealed that they belong to BVDV-1h group.

Validation of two commercial real-time RT-PCR kits for rapid and specific diagnosis of classical swine fever virus

2008 ◽  
Vol 147 (1) ◽  
pp. 136-142 ◽  
Author(s):  
M. Le Dimna ◽  
R. Vrancken ◽  
F. Koenen ◽  
S. Bougeard ◽  
A. Mesplède ◽  
...  
Keyword(s):  
Real Time ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Fever Virus ◽  
Rt Pcr ◽  
Specific Diagnosis

scholarly journals Analysis of hepatitis C virus/classical swine fever virus chimeric 5′NTRs: sequences within the hepatitis C virus IRES are required for viral RNA replication

2003 ◽  
Vol 84 (7) ◽  
pp. 1761-1769 ◽  
Author(s):  
Chantal B. E. M. Reusken ◽  
Tim J. Dalebout ◽  
Peter Eerligh ◽  
Peter J. Bredenbeek ◽  
Willy J. M. Spaan
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Classical Swine Fever Virus ◽  
Classical Swine Fever ◽  
Rna Replication ◽  
Fever Virus ◽  
Viral Rna
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