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A Rapid and Inexpensive Kit for On-Site Diagnosis of Vesicular Stomatitis and Food and Mouth Disease

Yilma, Tilahun
University of California - Davis
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  1. To develop chromatographic matrix strips coated with baculovirus-expressed VSV-N and FMDV-RdRp proteins and optimize the biochemical conditions and reaction sensitivity of such strips.
  2. To test the efficiency and accuracy of the diagnostic kit using serum and whole blood from known VSV-infected and non-infected naive cattle.
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NON-TECHNICAL SUMMARY: Vesicular stomatitis (VS) is an important disease of cattle, horses and pigs throughout the Americas. Food-and-mouth disease (FMD) is a highly contagious vesicular disease affecting up to 70 species of cloven-hoofed mammals, both domestic and wild. We would like to develop a rapid an dinexpensive penside diagnostic kit that can provide immediate on-site differentiation of VS from FMD.

APPROACH: The proposed diagnostic kit will determine the presence of anti-N or anti-RdRp antibodies in whole blood or serum samples. Serum samples will be obtained from known VSV-infected, FDMV-infected and uninfected calves (10/group). An internal control for the test will be incorporated in the final product to ensure the fidelity of the kit. Serum samples from the same animals will also be tested on the current VSV-specific and VMDV-specific diagnostic ELISA kits to evaluate accuracy.

PROGRESS: 2002/10 TO 2007/09
OUTPUTS: Foot and mouth disease and vesicular stomatitis cannot be distinguished clinically. Although vesicular stomatitis causes losses in animal productivity by itself, the biggest impact is the similarity to the truly devastating livestock disease, foot and mouth disease. Thus we have worked on a kit to rapidly diagnose and distinguish animals infected with the vesicular stomatitis virus from those infected with foot and mouth disease virus. Results from this project will be disseminated through presentations at national and international meetings or conferences and by publication in scientific journals.
PARTICIPANTS: Dr. Tilahun Yilma (DVM, PhD), VM:ILMB, UC Davis. Principal investigator. Dr. Shabbir Ahmad, developed recombinant baculovirus high-level expression viruses expressing the VSV N protein and purified the recombinant protein for the test kits. Dr. Leslie Jones, VM:ILMB, UC Davis, co-investigator, designed animal studies, immune assays, trained and supervised graduate students, postdocs, and technicians. Francisco Monge-Navarro (DVM), VM:ILMB, graduate student, developed the recombinant P3D protein. Shirley Leung, VM:ILMB, UC Davis, laboratory supervisor trained and supervised graduate students, postdocs, technicians, and lab assistants in lab safety. Ensured lab equipment maintenance, supplies, and compliance with university regulations. Sandy Yan, VM:ILMB, UC Davis, laboratory technician. This project provided training for graduate students and postdoctoral scholars.
TARGET AUDIENCES: Organizations and individuals involved in the control and prevention of reportable livestock diseases.

IMPACT: 2002/10 TO 2007/09
Vesicular Stomatitis (VS) is an economically important disease of livestock throughout the Americas. Clinical signs in cattle and pigs are undistinguishable from foot-and-mouth disease (FMD), one of the most devastating viral diseases of livestock. Currently, outbreaks of FMD-free areas must be diagnosed by laboratory methods and affected farms must be quarantined until laboratory results confirm whether it is VS, FMD, or another vesicular disease. To expedite the test and reduce expenses, we proposed to develop a rapid and simpler diagnostic tool which is needed to identify VS and FMD on site. The proposed diagnostic kit utilizes two different proteins from VSV and FMDV inexpensively produced in the baculovirus expression system to check for the presence of specific antibodies in animal sera. We have developed baculovirus expression systems producing large quantities of N protein of VSV. In addition, we have also developed a baculovirus expression system to produce a highly conserved recombinant FMDV protein (P3D) in large quantities. These baculovirus-produced antigens have been used to develop ELISA kits for detection of antibody. They have also been used to develop lateral flow chromatographic pen-side kits for detection of antibody to these two diseases. We are also developing monoclonal antibodies for development of similar pen-side kits to detect antigen in blood, sera or vesicular fluid.

Funding Source
Nat'l. Inst. of Food and Agriculture
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Legislation and Regulations
Bacterial Pathogens