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

• Published in: Journal of virological methods Vol. 92; no. 1; pp. 19 - 22
• Main Authors: Vilček, Štefan, Strojny, Ladislav, Ďurkovič, Branislav, Rossmanith, Wigbert, Paton, David
• Format: Journal Article
• Language: English
• Published: London Elsevier B.V 01.03.2001; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Biological and medical sciences; Bovine viral diarrhea virus; Bovine viral diarrhoea virus; Bovine Virus Diarrhea-Mucosal Disease - virology; BVDV; Cattle; Detection; Diarrhea Viruses, Bovine Viral - genetics; Diarrhea Viruses, Bovine Viral - isolation & purification; Filter; Fundamental and applied biological sciences. Psychology; Microbiology; Reverse Transcriptase Polymerase Chain Reaction; RNA, Viral - analysis; RT-PCR; Specimen Handling - methods; Specimen Handling - veterinary; BVDV; Bovine viral diarrhoea virus; Detection; Filter; RT-PCR; Pestivirus; Typing; Bovine; Diarrhea; Blood; Virus; Polymerase chain reaction; Vertebrata; Mammalia; Storage; Infectivity; Bovine diarrhea virus; Digestive diseases; Intestinal disease; Serum; Artiodactyla; Diagnosis; Flaviviridae; Ungulata
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/S0166-0934(00)00258-5

Of four solid carriers tested, Whatman paper No 1 was the best for storing blood and serum samples for the diagnosis of bovine viral diarrhoea (BVD) by means of viral RNA detection. The filter papers were impregnated with 10 μl of blood or serum, followed by air drying. Samples collected in this way from persistently infected animals had lost infectivity within a few days, but viral RNA could still be detected by RT-PCR for up to 6 months. When investigated by RT-PCR, 12 blood and 10 serum samples selected at random from animals persistently infected with BVD virus showed the same results whether samples had been spotted onto filters or examined directly from the liquid state. The filters spotted with blood or serum are convenient for storage and transport of samples to a diagnostic laboratory without the need for cooling. Sequencing of amplified RNA can be used subsequently for genetic typing.