Transcript AC Diagnostics, Inc.

Novel Immunocapture RT-PCR
Kits for Detection of Plant Viruses
Jun Q. Xia and Chunda Feng
AC Diagnostics, Inc., Fayetteville, AR
We Believe in Agri-Diagnostics!
(www.acdiainc.com)
Detection of Plant Viruses
1. Plant virus infections induce tremendous
economic losses.
2. Accurate and reliable detection of plant
viruses is critical.
3. Current ELISA detection methods may not
provide the sensitivity needed.
4. RT-PCR requires costly and time-consuming
viral sample RNA extraction.
Immunocapture RT-PCR
Coat antibody
PCR
tubes/plate on PCR tube
Immunocapture
incubation
Virus particle
captured by
antibody
Virus RNA
released from
particle
RT-PCR
amplification
♦ Combines two widely used detection technologies - ELISA and PCR.
♦ Eliminates costly and time-consuming sample RNA extraction procedure.
♦ Reduces risks from cross-contaminations in sample process .
♦ Shortens the test time and reduce assay cost for an accurate disease
diagnosis.
♦ Makes it possible to routinely conduct RT-PCR assays for large numbers of
plant samples.
Product Development
Antibody-Coated PCR tube:
• Coat antibody on PCR tube
• Post-coat and stabilize the coating antibody in PCR tube
Specific primers/probe:
• Develop the specific single-, duo-, multiple- or degenerate primers
• Design and label the probe with a fluorochrome and a quencher
Conventional or Real-time RT-PCR:
• Optimize the RT-PCR reaction system
• Validate the RT-PCR system for its sensitivity, specificity and
reliability.
Immunocapture RT-PCR Kit
☺Pre-coated PCR tubes for capturing virus particles
which can be directly amplified by RT-PCR
☺Optimized PCR primers which give bust and
reliable test results
☺Include all components and ready to use
☺Cost-effective and use-friendly, and being suitable
or PCR tests of large number of samples
Kit Formats
Immunocapture RT-PCR Kit
Immunocapture Kit
• 1X8 PCR strips pre-coated
with specific antibody
• PCR buffers - sample
extraction and washing
• Positive and negative
controls
• Instruction
• 1X8 PCR strips pre-coated
with specific antibody
• PCR buffers - sample
extraction and washing
• Positive and negative
controls
• RT-PCR primers,
• Master Mix (Optional)
• Reverse Transcriptase, and
Taq Polymerase (Optional)
• Instruction
Assay Procedure
of
Immunocapture
RT-PCR Kit
Immunocapture RT-PCR Kits Developed:
Alfalfa Mosaic Virus (AMV)
Potato Virus X (PVX)
Barley Stripe Mosaic Virus (BSMV)
Potato Virus Y (PVY)
Cherry Leaf Roll Virus (CLRV)
Tobacco Etch Virus (TEV)
Impatiens Necrotic Spot Virus (INSV)
Tobacco Mosaic Virus (TMV)
Maize Chlorotic Mottle Virus (MCMV)
Tobacco Ringspot Virus (TRSV)
Maize Dwarf Mosaic Virus (MDMV)
Tomato Aspermy Virus (TAV)
Pea Enation Mosaic Virus (PEMV)
Tomato Mosaic Virus (ToMV)
Peanut Stunt Virus (PSV)
Tomato Ringspot Virus (ToRSV)
Pepino Mosaic Virus (PepMV)
Wheat Streak Mosaic Virus (WSMV)
Pepper Mild Mottle Virus (PMMoV)
Immunocapture Kits:
Immunocapture kits are currently available for
detection of more than 50 plant viruses at AC
Diagnostics Inc.
Benefits by Using Immunocapture RT-PCR Kit
Getting accurate and consistent result, every
time running a RT-PCR.
 Saving funds, by capturing vial particles on
PCR tube without need of expensive RNA
extraction kits.
 Saving times, by eliminating the timeconsuming sample RNA extraction procedure.
 Flexible applications, by providing universal
PCR protocol to fit different PCR cycling
systems.
 High sensitivity, using for plant samples with
low pathogen titer such as seeds, woody plants
and stock plants.
Acknowledgments
Funding support provided by the USDA NIFA SBIR
Phase-I Grant project and now a Phase II Project
Novel Immunocapture RT-PCR Kits for
Detection of Plant Viruses
Jun Q. Xia and Chunda Feng
AC Diagnostics, Inc., Fayetteville, AR
We Believe in Agri-Diagnostics!
(www.acdiainc.com)