Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1998-05-21
2000-09-05
Fredman, Jeffrey
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 6, 435 912, 536 2432, 536 2433, C12Q 170, C12P 1934, C07H 2104
Patent
active
061141127
ABSTRACT:
The present invention provides new methods for preparing compositions for immunizing chickens against IBDV. The method comprises: amplifying RNA extracted from a chicken suspected of having IBDV by using a reverse transcriptase-polymerase chain reaction and a 3'end primer, referred to hereinafter as "Primer 1", which binds to a region of the negative strand of the VP2 gene of IBDV, said region extending from about nucleotide 2402 to about nucleotide 2463 on the negative strand, and a 5' end primer, referred to hereinafter as "Primer 2", which binds to a region of the positive strand of the VP2 gene, said region extending from about nucleotide 1424 to about nucleotide 1489 of the positive strand, to provide a PCR product of from about 733 to about 753 base pairs in length; digesting the PCR product with the restriction enzyme BstNI to provide BstNI restriction fragments; assaying the BstNI restriction fragments to determine the IBDV which has infected the chicken; isolating the IBDV from the chicken; treating the isolated IBDV to produce a killed virus or a live attenuated virus; and combining the killed virus or the live attenuated virus with a pharmacological carrier to provide a composition for immunizing chickens against infection from IBDV, particularly from infection with IBDV in the same class as the isolated virus. In a preferred embodiment, the method further comprises digesting an aliquot of the PCR product with the restriction enzyme MboI to provide MboI restriction fragments; and assaying the MboI restriction fragments to further classify the IBDV which has infected the chicken.
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Einsmann Juliet C.
Fredman Jeffrey
The Ohio State University
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