Specific, highly sensitive, nested PCR detection scheme for the

Details Specific, highly sensitive, nested PCR detection scheme for the Specific, highly sensitive, nested PCR detection scheme for the

1998-04-29

2000-05-30

MacMillan, Keith D.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving virus or bacteriophage

435 6, 435 4, C12Q 170

Patent

active

060689749

ABSTRACT:
The present invention provides for a highly sensitive nested polymerase chain reaction (PCR) method for detecting the presence or absence of the pseudorabies virus (PVR). The method targets a 674 base-pair region of the pseudorabies virus gII gene. Nucleotide sequences for highly specific novel primers derived from this gII region are also disclosed. These primers are used with the nested polymerase chain reaction method to amplify targeted nucleotide sequences within the 674 base-pair region of the gII gene. The novel primers and optimized reaction conditions of the nested polymerase chain reaction method enable significantly greater specificity for the viral DNA in tissue suspected of harboring the latent pseudorabies virus.

REFERENCES:
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patent: 5487969 (1996-01-01), Eberle et al.
patent: 5545523 (1996-08-01), Batt et al.
Robbins, et al., "The Pseudorabies Virus gII Gene Is Closely Related to the gB Glycoprotein Gene of Herpes Simplex Virus" Journal of Virology, vol. 61, No. 9, Sep. 1987, pp. 2691-2701.
Maes, et al., "Polymerase Chain Reaction Amplification of Pseudorabies Virus DNA from Acutely and Latently Infected Cells" Veterinary Microbiology, vol. 24, 1990, pp. 281-295.
Andrew K. Cheung, "Detection of the Large Latency Transcript of Pseudorabies Virus by RNA-PCR and Its Potential in Diagnosis" Journal of Veterinary Diagnostic Investigation, vol. 6, 1994, pp. 483-486.
Brown, et al., "Detection of Pseudorabies Viral DNA in Tonsillar Epithelial Cells of Latently Infected Pigs" American Journal of Veterinary Research, vol. 56, No. 5, May 1995, pp. 587-594.
Andrew K. Cheung, "Investigation of Pseudorabies Virus DNA and RNA in Trigeminal Ganglia and Tonsil Tissues of Latently Infected Swine" American Journal of Veterinary Research, vol. 56, No. 1, Jan. 1995, pp. 45-50.
Maes, et al., "Recent Developments in Latency and Recombination of Aujeszky's Disease (Psuedorabies) Virus" Veterinary Microbiology, vol. 55, 1997, pp. 13-27.
L.W. Enquist, PubMed Nucleotide Query, Accession No. M17321, Nov. 1987, pp. 1-3 (Computer-readable sequence), as published in Robbins, et al., "The Pseudorabis Virus gII Gene Is Closely Related to the gB Glycoprotein Gene of Herpes Simplex Virus" Journal of Virology, vol. 61, No. 9, Sep. 1987, pp. 2691-2701.

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