3. Research Priorities: Vector Area

• National Control System ?

• Policy/Plan
• Vector Control Measures
  • Community involvement
  • Insecticide management/resistance
    • Larvicide, adulticide
    • Biocontrol/predator
    • Medicinal plants
    • Personal protection
    • Ovitrap (lethal ovitrap)
• New Parameter
  • Stegomyia indices, BI, HI, CI
  • GIS, remote sensing
  • Nature of Mosquito Polymorphism
• Update on Enzyme Assay Screening
  • Due to constant evolution of insects (mosquitoes) which leads to isoenzyme/gene grouping
• Update on Biology Vector
  • Biting time, biting behavior
  • Population density
• Vector Competance
  • Genetic manipulation of vector
  • Identification of each vector as well as its susceptibility
• Transgenic from Vector to Non-vector
• Immune Response in Mosquitoes
  • Study of serine protease mediator
• Analysis of Salivary Enzyme
  • Allergies and reactions in man

4. Virology

Strain of Virus and the Severity of Infection

1. In vitro system

• Laboratory mutation
• NS₁ mutation

• Glycosylation
• Signal transduction
• Cell death and replication

• Capsid mutation

• Nuclear penetration
• Attenuation of replication
• Block encapsulation

• Role of NS₁ on complement (C’ ) activation

• Correlation of complement (C’ )activation and severity

• 5’ Non coding region (NCR) mutation

• Possibilities of vaccine candidates
• Propose new strategies for candidate testing – cytokine array

• Field isolates
  • 5’ NCR mutation
  • Natural attenuation

Pathogenesis

• Genomic and virulence of infection
  • Gene profile by micro-array analysis
  • Signature of DF and DHF
• Nitric Oxide (NO) in viral replication and pathogenesis
  • NO on molecular replication inhibition
  • Role of NO in DF and DHF
• T-cell immunity in DF and DHF patients

Future Research Plan

• Virology
  • Phylogenic analysis for Thai isolates and its relationship with severity
• Pathogenesis
  • Mechanisms of neutralization
  • Mechanism of apoptosis

• Destruction or protection
• Viral factors
• Host factor (serum factor)

• Effect of co infection on severity of illness?

5. Diagnostics

• Early

• Surrogate markers for prognosis of severity

Immunodiagnosis

1. Antibody detection

• Rapid test
• Immunochromatographics (recombinant E protein)
• ELISA IgM, IgG
  • Cost advantage

2. Antigen detection

• Predicting other factors like monoclonals to E, Pr, NS1
• NS1: 1-10 ng/ml

1-100 ng/ml

Hopes

• Early diagnosis
• Prediction of severity
• Understanding pathogenesis

Plans

• Evaluate clinical specimens
• Compare immunodiagnosis with molecular tests (NATS)

The issue of sharing monoclonal was brought up, to which Dr. Prida agreed.   
 

Molecular Tests (NAAT)

1. RT-nested PCR

• RT-single-PCR multiplex
• Microchip detection
• Qualitative
• Viral load (fluorescence & Chemiluminescence-based)

2. Real-time RT-PCR

• Quantitative
• Early diagnosis and surrogate marker of severity
• Costs:1000-1500 Baht

3. Detection of viruses in different compartments

6. Epidemiology

Direction:

• Burden
• Prediction (epidemiology and severity)
• Community database

7. Existing Infrastructure and problems
1. Epidemiology data

• National surveillance form 506

• Serotyping surveillance

2. Problems

• Coverage
• Quality
• Validity
• Accessibility