The meningococcus

1. The meningococcus Nathan Keller Md. PhD. The Department of clinical microbiology and the national center for meningococci The Chaim Sheba Medical Center Tel Hashomer

2. Meningococcal Disease Etiologic Agent: • Neisseria meningitidis (Gram-negative diplococcus bacterium) with multiple serogroups ( A, B, C, D, 29E, H, I, K, L, W-135, X, Y, and Z). • Strains belonging to groups A, B, C, Y and W-135 are implicated most frequently in invasive disease.

3. Neisseria gonorrhoeae Gram negative intracellular diplococci

4. The meningococcus

5. Neisseria meningitidis • Aerobic gram-negative bacteria • At least 13 serogroups based on characteristics of the polysaccharide capsule • Most invasive disease caused by serogroups A, B, C, Y, and W-135 • Relative importance of serogroups depends on geographic location and other factors (e.g. age)

6. N. meningitidis(the "meningococcus")

7. Meningococcal meninigitis • second most common meningitis • pneumococcus, most common • fatal if untreated • responds well to antibiotic therapy • penicillin

9. N. meningitidis • resides in man only • usually sporadic cases • mostly young children • outbreaks • adults • crowded conditions • e.g. army barracks

10. Neisseriameningitidis • upper respiratory tractinfection • adhesion pili • bloodstream brain

11. Microbial routes to meninges

12. Capsule • capsule • inhibit phagocytosis • anti-capsular antibodies • stop infection • antigenic variation • serogroups • vaccine • multiple serogroups

13. Identification of infectious agents in the diagnostic laboratory • Aids treatment • Helps antibiotic selection • General hospital laboratory • physiological tests • Reference laboratories • Genetic tests

14. Laboratory Diagnosis • spinal fluid • Gram negative diplococci • within polymorphonuclear cells • meningococcal antigens • Culture • Thayer Martin agar • Choclate agar, Blood agar

15. Microscopy • spinal fluids (meningitis) • sensitivity poor

16. CSF- N. meningitidis

17. Isolation and identification • Step 2. Colonies Gram stained • cells observed microscopically

19. Step 3.Isolated bacteria are speciated • Generally using physiological tests

20. Typical Culture Laboratory Bench

21. Antibiotic susceptibility testing Susceptible Not susceptible Bacterial lawn Growth No growth Antibiotic disk Data on Usraeli susceptibility will be given by dr block

22. Neisseria meningitidesDiagnosis • Immediate • Microscopy • Antigen detection • Molecular

24. Neisseria in Leukocytes

25. Meningococcemia - intracellular diplococci in buffy coat leukocytes

26. Neisseria meningitidesDiagnosis • Antigen detection

27. Antigen detection assay Kurzynski et al JCM jun 1985

28. Real-time PCR ds DNA Cycle one Dye Cycle two Cycle 30 2 30

29. DNA-DNA hybridization Strain 1 Heat + Strain 2 0% Homology 100% Homology

30. Cell surface of Neisseria meningitidis Capsule - polysacch. serogroup A, B, C, Y, W-135, etc. Pilus Outer membrane phospholipid Opa (opacity - protein) porin (serotype 2,15,etc.) lipooligosaccharide Peptidoglycan layer Cytoplasmic membrane Cytoplasm

31. Case Definition Clinical Description: Meningococcal disease manifests most commonly as meningitis and/or meningococcemia that may progress rapidly to purpura fulminans, shock, and death. However, other manifestations might be observed. Laboratory criteria for diagnosis: Isolation of Neisseria meningitidis from a normally sterile site (e.g., blood or cerebrospinal fluid (CSF) or, less commonly, joint, pleural, or pericardial fluid)

32. Case Definition Case Classification Probable:a case with a positive antigen test in cerebrospinal fluid or clinical purpura fulminans in the absence of a positive blood culture. What about real time PCR? Confirmed:a clinically compatible case that is laboratory confirmed.

33. באדיבותה של ד"ר אמיליה אניס המחלקה לאפידמיולוגיה משרד הבריאות

34. Laboratory confirmed isolates of Neisseria meningitidis from blood and cerebrospinal fluid 1986 - March 2005

35. Laboratory confirmed isolates of Neisseria meningitidis from blood and cerebrospinal fluid – 1986 - March 2005

36. Laboratory confirmed isolates of Neisseria meningitidis from blood and cerebrospinal fluid – 1986 - March 2005

37. Laboratory confirmed isolates of Neisseria meningitidis from blood and cerebrospinal fluid – 1986 - March 2005

38. Laboratory confirmed isolates of Neisseria meningitidis from blood and cerebrospinal fluid – 1986 - March 2005

39. Meningococci this year • Gr B 21 (32) • Gr Y 7 (8) • Gr W135 2 • Polyagglut 1 (1) • Gr A - (1) • Gr C - (4) • (Number of cases in 2004)

40. λ M11 M13 M18 M19 M20 M21 λ M23 M24 M25 M26 M28 λ בידודי חיידקי מנינגוקוקים שהתקבלו לפי מקורם: PFGE of GR B meningococci Feb-Mar 2005 λ – סמן גודל

41. Dice (Tol 1.0%-1.0%) (H>0.0% S>0.0%) [0.0%-100.0%] PFGE NheI No. Hospital Residence % 100 40 60 80 M20 . Hadassa ein karem Beit Shemesh M28 . Ichilov Tira M11 . Belinson Kiriat Sefer M26 . Sheba Ashdod Belinson Ramat Gan M19 . M13 Kaplan Yavne M18 Sheba Bnei Brak M25 Kaplan Ashdod M21 Volfson Holon Sheba Holon M23 Volfson Holon M24 PFGE analysis of GR B meningococci באדיבותה של ד"ר לאה ולינסקי המעבדה לביולוגיה מולקולרית מעבדות מרכזיות משרד הבריאות ירושלים

42. Neisseria gonorrhoeae Gram negative intracellular diplococci

44. תודות • ד"ר לאה ולינסקי • יהודית דוידסון • ד"ר גיל סמולן • פרידה יעקובוביץ • ד"ר גנריאטה רוזל • גיה סרקיסיאן • ד"ר בלוק • צוות המעבדה