Detection of extraneous agents in vaccines using the polymerase chain reaction of Newcastle disease virus in poultry biologicals
Bruckner, L.; Stäuber, N.; Brechtbühl, K.; Hofmann, M.A.
Developments in Biological Standardization 86: 175-182
1996
ISSN/ISBN: 0301-5149 PMID: 8785947 Document Number: 456702
Reverse transcription-polymerase chain reaction (RT-PCR) in poultry vaccine was applied to the detection of Newcastle disease virus (NDV), using two primer pairs spanning the cleavage site of the FO fusion protein coding sequence. Amplification of a specific cDNA segment was possible from live and inactivated, oil-adjuvanted NDV vaccines without previous treatment. The RT-PCR was able to detect between 5 x 10(2) EID50 (in live vaccine preparations) and 10(5) EID(50) or 0.056 haemagglutinating units of NDV (in inactivated vaccine preparations). In addition, live vaccine preparations were inactivated with beta-propiolactone (beta-PL). Amplified cDNA was obtained after treatment with 0.1% beta-PL, whereas at a concentration of 1% or 10% no specific bands were visible in the agarose gel. These results demonstrate the applicability of the method for the control of poultry vaccines by ensuring the absence of extraneous agents.