Assessment of inactivation of hepatitis A vaccine by compound PCR.
Assuring the complete inactivation of hepatitis A virus (HAV) vaccine commonly requires prolonged tissue culture amplification, followed by detection of virus antigen in cell lysates. A reliable, but faster, alternative procedure is highly desirable since it will permit the prescreening of experimental batches of killed HAV, prior to tissue-culture amplification. We established experimental conditions for simultaneous, polymerase chain reaction (PCR)-based amplification of viral and cellular mRNA sequences from infected cell RNA (compound PCR). Under these conditions, the presence of virus-specific amplified sequences, as detected by Southern blot, allows the identification of incompletely inactivated vaccine batches with a threshold practically identical to that of the more time-consuming subculture and ELISA. Compound PCR is, by its nature flexible enough for adaption to different requirements and it should prove useful for rapid prescreening of vaccine batches and pilot studies for improvement of inactivation protocols.
Fineschi
N
N
Molecular Virology, I.R.I.S. Srl., Siena, Italy.
Pellegrini
V
V
Zuckerman
A J
AJ
Cavalieri
F
F
D001483:Base Sequence; D015342:DNA Probes; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D022362:Hepatitis A Vaccines; D006507:Hepatovirus / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D015164:Vaccines, Inactivated / Q000302:isolation & purification; D014761:Viral Hepatitis Vaccines / Q000302:isolation & purification*; D014773:Virology / Q000379:methods
D015342:DNA Probes; D022362:Hepatitis A Vaccines; D012367:RNA, Viral; D015164:Vaccines, Inactivated; D014761:Viral Hepatitis Vaccines