Method and kit for detecting white spot syndrome virus

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C435S091100, C435S091200, C536S023100, C536S024330, C536S025320, C536S026600

Reexamination Certificate

active

06872532

ABSTRACT:
A method for detecting white spot syndrome virus (WSSV) in a sample includes the steps of (a) adding to the sample a thermostable polymerase, appropriate nucleoside triphosphates, a nucleic-acid-binding fluorescent entity, and a pair of primers substantially complementary to a target nucleic acid having the sequence shown in SEQ ID NO: 1 or the complement of the target nucleic acid; (b) thermally cycling the sample between at least a denaturation temperature and an elongation temperature; (c) illuminating the sample with a selected wavelength of light that is absorbed by the fluorescent entity during the thermally cycling step; (d) determining the amount of fluorescence generated by the fluorescent entity; and (e) detecting the presence of the target nucleic acid by analyzing the amount of luminescence determined after at least one amplification cycle. The invention further provides kits for detection of WSSV in samples.

REFERENCES:
patent: 5824535 (1998-10-01), Kou et al.
patent: 6190862 (2001-02-01), Kou et al.
Arun K. Dhar et al., “Detection and Quantification of Infectious Hypodermal and Hematopoietic Necrosis Virus and White Spot Virus in Shrimp Using Real-Time Quantitative PCR and SYBR Green Chemistry”, Journal of Clinical Microbiology, Aug. 2001, pp. 2835-2845.
Belcher CR et al., “Colourimetric PCR-based Detection of Monodon Baculovirus in Whole Penaeus Monodon Postlarvae”, J. Virol. Methods, 1998 Sep.; 74(1):21-9 (Abstract Only).
Nunan LM et al., “Development of a Non-Radioactive Gene Probe by PCR for Detection on White Spot Syndrome Virus (WSSV)”, J. Virol. Methods, Jan;63(1-2):193-201 (Abstract Only).
Hsu HC et al., “Studies on Effective PCR Screening Strategies for White Spot Syndrome Virus (WSSV) Detection in Penaeus Mondon Brooders”, Dis. Aquat. Organ., 1999 Dec. 22;39(1):13-9.
Venegas CA et al., “Quasi-Immune Response of Penaeus Japonicus to Penaeid Rod-Shaped DNA Virus (PRDV)”,0 Dis. Aquat. Organ., Aug. 31, 2000;42(2):83-9 (Abstract only).
Tang KF et al. “Detection and Quantification of Infectious Hypodermal and Hematopoietic Necrosis Virus in Penaeid Shrimp by Real-Time PCR”, Dis. Aquat. Organ., Mar. 9;44(2):79-85 (Abstract Only).
Dhar AK et al., “Detection and Quantification of Infectious Hypodermal and Hematopoietic Necrosis Virus and White Spot Virus in Shrimp Using Real-Time Quantitative PCR and SYBR Green Chemistry”, J. Clin. Microbiol., Aug. 2001;39(8):2835-45 (Abstract Only).
Abstract and Machine Translation of JP9201196 (published Aug. 5, 1997).

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