Detection of extraneous agents in vaccines using the polymerase chain reaction of Newcastle disease virus in poultry biologicals.
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.
Bruckner
L
L
Institute of Virology and Immunoprophylaxis, Mittelhäusern, Switzerland.
Stäuber
N
N
Brechtbühl
K
K
Hofmann
M A
MA
D000826:Animal Testing Alternatives / Q000379:methods*; D000818:Animals; D001483:Base Sequence; D001688:Biological Products / Q000302:isolation & purification*; D002645:Chickens; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004340:Drug Contamination; D008969:Molecular Sequence Data; D009522:Newcastle disease virus / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D011200:Poultry / Q000276:immunology* / Q000821:virology*; D012680:Sensitivity and Specificity; D014612:Vaccines / Q000302:isolation & purification
D001688:Biological Products; D017931:DNA Primers; D004279:DNA, Viral; D014612:Vaccines