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GENUS: NEISSERIA

GENUS: NEISSERIA. Amany Niazy 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. SPECIMEN.

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GENUS: NEISSERIA

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  1. GENUS: NEISSERIA Amany Niazy Lab # 6

  2. 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.

  3. SPECIMEN • Neisseria gonorrhoea Urethral discharge  Genital swabs (servical or rectal) • Neisseria meningitidis CSF • Fastidious organism very sensitive to environmental factors Process specimen as soon as possible

  4. GRAM STAIN • Direct Gram Stain: • Neisseria are intracellular • organisms • GNDC inside and outside pus cells (PMN) (kidney or bean shaped)

  5. CULTURE • Inoculate into • Chocolate Agar • Basal media  at 40C (no solidification) add sheep blood • Keep in water bath at 70C to hemolyse blood • This releases and denatures many RBCs and serum proteins, • making them more readily digestible to the bacteria • Colonies appear: Colorless , raised, 1-2 mm in diameter

  6. CULTURE • Thayer Martin (selective medium) • Contain antibiotics to inhibit other bacteria and fungi VCNT Vancomycin, Colistin, Nystatin, Trimethoprim.

  7. INCUBATION • Temp 35-37 ˚C ONLY • Need 5-10% CO2 to grow: • CO2 envelop in anaerobic jar or • Candle jar • For 48 hours Non-pathogenic Neisseria grow at room temperature

  8. IDENTIFICATION • Gram stain  GNDC • Oxidase +ve • Carbohydrate utilization test

  9. IDENTIFICATION • Gram stain from plate  GNDC (Kidney shaped)

  10. IDENTIFICATION • All Neisseria species are Oxidase +ve • Preparation of Oxidase Test: • Add filter paper on a clean slide • Add 2-3 drops of oxidase reagent on filter paper • Use cover slip to transfer colony onto filter paper • + ve Purple color

  11. IDENTIFICATION • HISS medium. broth + serum + sugar + indicator (phenol red) Neisseria meningitides Neisseria gonorrhoeae

  12. YOUR WORK FOR TODAY • Prepare 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: • Immunoassays • DNA hybridization • PCR • Doc Kaiser’s microbiology website http://student.ccbcmd.edu/~gkaiser/goshp.htm

  14. Numbers in Saudi Population 13 cases of meningococcal meningitis in 2007

  15. THANK YOU

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