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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. Plant virus infections induce tremendous economic losses .

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slide1

Novel Immunocapture RT-PCR Kits for Detection of Plant Viruses

Jun Q. Xiaand Chunda Feng

AC Diagnostics, Inc., Fayetteville, AR

We Believe in Agri-Diagnostics!

(www.acdiainc.com)

slide2

Detection of Plant Viruses

  • Plant virus infections induce tremendous economic losses.
  • Accurate and reliable detection of plant viruses is critical.
  • Current ELISA detection methods may not provide the sensitivity needed.
  • RT-PCR requires costly and time-consuming viral sample RNA extraction.
slide3

Immunocapture RT-PCR

  • Virus particle captured by antibody
  • Virus RNA released from particle
  • RT-PCR amplification
  • Immunocapture incubation
  • Coat antibody on PCR tube

PCR tubes/plate

  • ♦ 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.
slide4

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.
slide5

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
Kit Formats

Immunocapture Kit

Immunocapture RT-PCR Kit

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

  • 1X8 PCR strips pre-coated with specific antibody
  • PCR buffers - sample extraction and washing
  • Positive and negative controls
  • Instruction
slide7

Assay Procedure

of Immunocapture RT-PCR Kit

slide8

Immunocapture RT-PCR Kits Developed:

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
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 time-consuming 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.
slide10

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. Xiaand Chunda Feng

AC Diagnostics, Inc., Fayetteville, AR

We Believe in Agri-Diagnostics!

(www.acdiainc.com)