EBPL Training Program Curriculum

Training For PCR Diagnosis Of Shrimp Virus

1. Introduction to molecular biology and basic concept of PCR

• DNA structure
• Components of DNA
• The function of DNA polymerase
• The principle of polymerase chain reaction (PCR)
• Basic requirements and design of PCR lab
• Potential problems of PCR

2. Introduction of PCR related equipment

• DNA extraction kit
• IQ2000 WSSV Kit
• Reagents for electrophoresis

3. Major shrimp Viruses and Viral diseases

• MBV,HPV, IHHNV, & WSSV
• White spot syndrome virus (WSSV).
• Yellow-head virus/Gill-associated virus (YHV/GAV)

4. Prevention of Viral Diseases.

• Avoid vertical transmission
• Avoid horizontal transmission

5. Practice of DNA extraction

• Crude DNA Extraction
• Pure DNA Extraction

6. Practice of PCR operation procedure

• DNA extraction kit
• IQ2000 WSSV Kit
• Reagents for electrophoresis

7.  Practice of gel electrophoresis

• Agarose gel preparation
• Electrophoresis
• Gel staining and data assay

8. Sampling procedure and sample preservation

• Preservation of shrimp (tissue) samples
• Preservation of DNA samples
• Introduction of sample dissecting tools
• Disinfection procedure of the sample dissecting tools
• Sampling strategy for brood stocks
• Sampling strategy for eggs and nauplii
• Sampling strategy for PLs
• Sampling strategy for juveniles in the grow-out pond
• Sampling strategy for environmental factors

9. Perform the assay by using real samples

• To compare the sensitivity, reproducibility, and to practice more
• Positive standards and negative control are required.
• Requirements of sample

10. Detail description of PCR diagnostic procedure

• Sampling, sensitivity, and statistical issue
• False positive or false negative result
• Reproducibility
• Lab management

11. How to integrate the diagnostic results into practical shrimp farming

• Environmental screening
• Choosing the source of brooders and PLs
• Viral risk management for the grow-out pond

• Real samples operation
• Discussion