Annexes to the Laboratory Manual Homepage
The annexes provide basic test method details or guidance documents to augment the material in the laboratory manual. The annex materials and protocols are not intended to provide laboratories with a standard operating protocol (SOP) as is, but may serve as a template for each individual laboratory to develop SOPs, and to stay current with new techniques as appropriate according to availability and resources.
The inclusion of reagents and products from specific manufacturers does not constitute an endorsement by the WHO or the GMRLN.
The documents are organized by subject matter into groups designated by a letter (A-F) and a number. Each annex in the list is assigned a consecutive number. If a document is superseded by a revised version, the new version will have a numeric extension. For example, if a revised version of an existing document named Annex B5 becomes available, the revised document will replace the original document and will be referred to as Annex B5.2.
New protocols will be added to the appropriate group with the next consecutive number at the end of the existing list.
The annex groups are organized according to subject matter described below. Click on the annex group below to navigate to the page with the directory (list) of the individual protocols (and links to pdfs) within the annex group.
Group A Annex Directory: Specimen collection, transport, and processing
- Annex A1. Measles and rubella laboratory request form
- Annex A2. Measles and rubella investigation form
- Annex A3. Shipping clinical specimens
- Annex A4. Dried blood spot (DBS) protocols [Incomplete]
- Annex A5. Processing oral fluid samples and evaluation of total IgG [Pending]
- Annex A6. Preparation and use of FTA cards
Group B Annex Directory: Protocols and documents for specimen testing (case confirmation)
- Annex B1. Comparison of measles and rubella IgM kits [placeholder document, pending update]
- Annex B2. EuroImmun protocol for measles IgM ELISA - English | Annex B2 - French
- Annex B3. EuroImmun protocol for rubella IgM ELISA - English | Annex B3 - French
- Job aid for Annex B3 - see Annex B2a ( English & French versions)
- Worksheet (manual calculation, EuroImmun) for Annex B3 – see Annex B2b
- Worksheet (built-in calculations, EuroImmun) for Annex B3 – see Annex B2c
- Annex B4. Serion protocol for measles IgM ELISA - English | Annex B4 - French
- Annex B5. Serion protocol for rubella IgM ELISA - English | Annex B5 - French
- Job aid for Annex B5 - see Annex B4a (English & French versions)
- Worksheet (manual calculation, Serion) for Annex B5 - see Annex B4b
- Worksheet (built-in calculations, Serion) for Annex B5 – see Annex B4c Annex B6.
- Annex B6. Testing oral fluid for measles- or rubella-specific IgM by EIA [Pending]
- Annex B7. Propagation and maintenance of Vero/hSLAM cell culture (CDC)
- Annex B8. Measles and rubella virus isolation in cell culture [Pending]
- Annex B9. Confirmation of rubella virus in culture by IFA and ICA
- Annex B10. RNA extraction methods for clinical samples and FTA cards
- Annex B11. Measles real-time RT-PCR, N gene (CDC)
- Annex B11a. Worksheet (duplicate wells) for Annex B11
- Annex B11b. Plate layout (duplicate wells) for Annex B11
- Annex B11c. Worksheet (triplicate wells) for Annex B11
- Annex B11.2. Measles real-time RT-PCR protocol
- Annex B11.2a. Invitrogen Real Time PCR Template
- Annex B12. Rubella real-time RT-PCR (NSP)
- Annex B13. Clean-up of PCR products
Group C Annex Directory: Protocols and documents for sequencing and genetic analysis
- Annex C1. Measles genotyping RT-PCR (CDC)
- Annex C2. Protocol for rubella genotyping RT-PCR, Qiagen (CDC)
- Annex C3. Protocol for rubella genotyping RT-PCR, Invitrogen Superscript III (CDC)
- Annex C3a. Worksheet for Annex C3 included in Annex C2a docx
- Annex C4. Measles sequencing protocol for genotyping (CDC)
- Annex C5. Rubella sequencing protocol for genotyping (CDC)
- Annex C6. Measles sequence analysis for genotyping using MEGA v6.06
- Annex C7. Rubella sequence analysis for genotyping using MEGA v5.05
- Annex C8. Measles & rubella sequence assembly (editing) using GeneStudio
- Annex C9. Phylogenetic analysis (MEGA v7 or X) of measles and rubella sequences assembled using Genestudio
- Annex C10. Agarose gel electrophoresis with measles and rubella synthetic positive controls
- Annex C11. Instructions for MeaNS [Pending]
- Annex C12. Instructions for RubeNS [Pending]
- Annex C13. Directions for finding and using the MeaNS unique sequence ID [Pending]
- Annex C14. Epidemiological charts [Pending]
Group D Annex Directory: Resources and information related to laboratory quality assessment or management
- Annex D1. Levey–Jennings control charts
- Annex D2. Supplemental information for optimal laboratory design
- Annex D3. Trouble shooting guide for IgM EIAs [Pending]
- Annex D4. Recommendations for the evaluation of EIAs for IgM detection [Pending]
- Annex D5. Verification for new IgM EIAs [Pending]
- Annex D6. Accreditation checklists [Pending]:
- Annex D6 - Part 1: Accreditation checklist Section 1 GENERAL
- Annex D6 - Part 2: Accreditation checklist Section 2 SEROLOGY
- Annex D6 - Part 3: Accreditation checklist Section 3 MOLECULAR
- Annex D6 - Part 4: Accreditation checklist Section 4 VIRUS ISOLATION
Group E Annex Directory: Techniques for additional characterization of specimens and cases
- Annex E1. Real-time RT-PCR for measles vaccine strains
- Annex E2. Measurement of rubella neutralizing antibody titer (rubella immunocolorimetric assay, CDC) [Pending]
- Annex E2a. Worksheet for Annex E2 [Pending]
- Annex E3. Plaque reduction neutralization assay for measles [Pending]
- Annex E4. Measles IgG avidity testing [Pending] Annex E5. Rubella IgG avidity testing [Pending]
- Annex E6. Extended sequencing (MF-NCR sequencing, CDC) [Pending]
- Annex E7. Protocol for measles genotyping with M13 primers (CDC)
- Annex E8. Nested RT-PCR for measles
- Annex E9. Nested RT-PCR for rubella [Pending]
Group F Annex Directory: Methods for measurement of population immunity
- Annex F1. Mixture modeling [Pending]
- Annex F2. Limitations and recommendations for measurement of measles and rubella IgG [Pending]
For inquiries, please contact:
Dr. Mick N. Mulders, Scientist
Coordinator Global VPD Laboratory Networks
Expanded Programme on Immunization (EPI)
Department of Immunization, Vaccines & Biologicals (IVB)
Universal Health Coverage/Lifecourse Division (UHC/LC)
Mail: muldersm@who.int