3. SEROLOGY
• I did the following procedures:
• Hemagglutination
• Hemagluttination Inhibition
4. HEMAGGLUTINATION
• Take normal saline 25 micro litre
• Add virus 25 micro litre
• Add blood 25 micro litre
• Note button formation for positive reults and calculate titer
• If HA result was 7 then titre was 16 ml normal saline in 0.25 ml virus
5. HEMAGLUTTINATION INHIBITION
• Take normal saline 25 micro litre
• Add serum 25 micro litre
• Add prepared virus 25 micro litre
• Refrigerate 30 min
• Add blood 25 micro litre
• Refrigerate 30 min
• Note the number of well with don’t have button formation
6. BACTERIOLOGY
• I did following procedures:
• Media preparation
• Culture
• Antibiotic sensitivity test
• Gram staining
7. MEDIA PREPERATION
• Take 1 litre water
• Add eosin methylene blue agar powder
• Autoclave for 15 min at 121 degree at 15 psi pressure
• Pour in petri dishes
8. CULTURE
• Take agar plate
• Take culture with wire loop from broth solution
• Add in agar plate with streaking
• Cover it
• Put in incubator for 24 hours
9. ANTIBIOTIC SENSITIVITY
• Take nutrient agar plate
• Add culture with normal saline with lawn technique
• Add anti biotic buttons
• Refrigerate for 24 hours
• Note the zone
10. GRAM STAINING
• Take heat fixed slide
• Add crystal violet stain for 1 min
• Add iodine for 30 sec
• Add alcohol for 110 sec
• Add saffrenin for 30 sec
• Examine under microscope at 100x with cedar wood oil
11. VIRUS ISOLATION AND HARVESTING
• I did these procedures:
• Virus isolation
• Virus harvesting
12. VIRUS ISOLATION
• Take normal saline
• Add in epindorf tubes
• Make 10 fold dilution of 0.1 ml virus
• Discard the first 5
• Take the rest 5
• Add them in fertile eggs by punching holes
• Observe every 24 hours with candling
• The dead eggs are refrigerated
• Rest are again sent to incubator
13. • Repeat 3 time
• After 72 hours refrigerate the eggs for 24 hours
14. VIRUS HARVESTING
• Break the eggs open
• Take allontic fluid and perform rapid HA test
• The eggs that have a very high number of virus are reported
• The rest of fluid is collected in a collection jar
