Global Alert and Response (GAR)

Laboratory methods for the diagnosis of meningitis caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae

WHO/CDS/CSR/EDC/99/7/EN


Laboratory Methods for the Diagnosis of Meningitis Caused by Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae

Introduction
Acknowledgements
I. Epidemiology of Meningitis Caused by Neisseria meningitidis, Haemophilus influenzae and Streptococcus pneumoniae
II. General Considerations
     A. Record Keeping
III. Collection and Transport of Clinical Specimens
      A.Collection of Cerebrospinal Fluid (CSF)
                      A1. Lumbar Puncture
      B. Collection of Blood
                      B1. Precautions
                      B2. Sensitivity of Blood Cultures
                      B3. Venipuncture
      C. Transport of Clinical Specimens
                     C1. CSF
                     C2. Blood                         
IV. Primary Culture, Subculture and Presumptive Identification
      A. Inoculation of Primary Culture Media
                     A1. CSF
                          1.1. Gram Stain Procedure for CSR (Hucker Modification)
                          1.2. General Methods for Performing Latex Agglutination Tests
                     A2. Blood
      B. Subculture
                    B1. Blood Culture Bottle
                    B2. T-1 Medium
      C. Macroscopic Examination of Colonies          
V. Identification of N. meningitidis
     
A. Kovac's Oxidase Test
      B. Identification of the N. meningitidis Serogroup
      C. Carbohydrate Utilization by N. meningitidis - Cystine Trypticase Agar Method
      D. Commercial Identification Kits
VI. Identification of S. pneumoniae
     
A. Susceptibility to Optochin
      B. Bile Solubility Test
      C. Slide Agglutination Test
VII. Identification of H. influenzae
     
A. Identification of the H. influenzae Serotype
      B. Identification of X and V Factor Requirements
                    B1. X, V and XV Paper Disks or Strips
                    B2. Haemophilus ID "Quad" Plates
VIII. Preservation and Transport of N. meningitidis, S. pneumoniae, and H. influenzae
      A. Short-Term Storage
      B. Long-Term Storage
                    B1. Preservation by Lyophilization
                    B2. Preservation by Freezing
      C. Transportation of Cultures
                    C1. Transport in Silica Gel Packages 
IX. Suggested Further Reading
X. Annexes
     A. Basic Requirements, Supplies and Equipment
     B. Media and Reagents
     C. Preparation of Media and Reagents
     D. Serotyping of S. pneumoniae 
     E. Biotyping of H. influenzae

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