Lab 6 NEISSERIA SPECIES Contains many species 2 are human pathogens Neisseria gonorrhoea gonococcus Neisseria meningitidis meningococcus Other species are normal flora that inhabit the upper respiratory tract of humans ID: 1009733
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1. GENUS: NEISSERIA Amany Niazy Lab # 6
2. NEISSERIA SPECIESContains many species; 2 are human pathogensNeisseria gonorrhoea (gonococcus)Neisseria meningitidis (meningococcus)Other species are normal flora that inhabit the upper respiratory tract of humans.
3. SPECIMENNeisseria gonorrhoea Urethral discharge Genital swabs (servical or rectal)Neisseria meningitidis CSFFastidious organism very sensitive to environmental factors Process specimen as soon as possible
4. GRAM STAINDirect Gram Stain:Neisseria are intracellular organismsGNDC inside and outside pus cells (PMN) (kidney or bean shaped)
5. CULTUREInoculate into Chocolate Agar Basal media at 40C (no solidification) add sheep bloodKeep in water bath at 70C to hemolyse bloodThis releases and denatures many RBCs and serum proteins, making them more readily digestible to the bacteriaColonies appear: Colorless , raised, 1-2 mm in diameter
6. CULTUREThayer Martin (selective medium)Contain antibiotics to inhibit other bacteria and fungiVCNT Vancomycin, Colistin, Nystatin, Trimethoprim.
7. INCUBATIONTemp 35-37 ˚C ONLY Need 5-10% CO2 to grow:CO2 envelop in anaerobic jar orCandle jarFor 48 hours Non-pathogenic Neisseria grow at room temperature
8. IDENTIFICATIONGram stain GNDCOxidase +veCarbohydrate utilization test
9. IDENTIFICATION Gram stain from plate GNDC (Kidney shaped)
10. IDENTIFICATIONAll Neisseria species are Oxidase +vePreparation of Oxidase Test: Add filter paper on a clean slideAdd 2-3 drops of oxidase reagent on filter paperUse cover slip to transfer colony onto filter paper+ ve Purple color
11. IDENTIFICATIONHISS medium.broth + serum + sugar + indicator (phenol red)Neisseria gonorrhoeaeNeisseria meningitides
12. YOUR WORK FOR TODAYPrepare gram stain from culture.Do oxidase test.Subculture organism to chocolate agar.Incubate in the CO2 jar.Read after 48 hours.See sugar utilization tube (demonstration) See intracellular GNDC slide (demonstration slide)
13. New diagnosis techniques:ImmunoassaysDNA hybridizationPCRDoc Kaiser’s microbiology websitehttp://student.ccbcmd.edu/~gkaiser/goshp.htm
14. Numbers in Saudi Population13 cases of meningococcal meningitis in 2007
15. THANK YOU