Standard operating procedures (SOPs)
The following APNET-approved standard operating procedures (SOPs) for specimen collection and laboratory diagnosis of EV71 infection are available for downloading as pdf files.
Specimen collection
Laboratory methods
The laboratory methods used by the APNET collaborators are based on those developed at the Institute for Health and Community Medicine, UNIMAS, Sarawak Malaysia and are taught at regular APNET workshops.
- SOP COVERSHEET
- CHART 1 - Specimen Processing & Virus Isolation Flowchart
- CHART 2 - PCR for Primary Specimens Flowchart
- CHART 3 - PCR for Virus Isolates Flowchart
- IKPK 001 - SPECIMEN COLLECTION/PROCESSING
- IKPK 002 - SPECIMEN ALIQUOTING
- IKPK 003 - RNA/DNA EXTRACTION
- IKPK 004 - PAN-ENTEROVIRUS RT-PCR FOR PRESENCE OF ENTEROVIRUS GENOME (PRIMERS MD90/MD91 IN 5'UTR)
- IKPK 006 - DNA EXTRACTION FROM AGAROSE GELS FOR SEQUENCING
- IKPK 007a - SEQUENCING REACTIONS - BigDye Terminator v3.0 sequencing Kit (Applied Biosystems)
- IKPK 007b - SEQUENCING REACTIONS - DYEnamic ET Dye Kit (Amersham)
- IKPK 008 - TISSUE CULTURE METHODS
- IKPK 009 - ENTEROVIRUS SEROTYPING VP4-BASED RT-PCR (PRIMERS EVP2/0L68-1)
- IKPK 010 - EV71-SPECIFIC VP1-BASED RT-PCR (PRIMERS MAS01S/MAS02A0
- REFERENCES
- Indirect immunofluorescence assay (IFA) for rapid, presumptive identification of EV71 in cell culture