SPADA Book - April 11, 2017

DRAFT ENVIRONMENTAL ORGANISMS PANEL, VERSION 4

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2.3 73 74 Inclusion of all environmental panel organisms is not a requirement if a method developer provides 75 appropriate justification that the intended use of the assay permits the exclusion of specific panel 76 organisms. Justification for exclusion of any environmental panel organism(s) must be documented and 77 submitted. 78 79 If bioinformatic analysis is completed then DNA from Group 1 organisms should be tested. If bioinformatic 80 analysis is not completed then DNA from Group 1 and 2 organisms should be tested. 81 82 Organisms may be tested as pools containing ten organisms each represented at 10 times the AMDL. Cell 83 lines should be tested individually at a DNA concentration equivalent to 10X AMDL. These tests should 84 be negative for the target sequence. The same pools can be spiked with 2X the AMDL for the target 85 organism, mixed and tested. These tests should be positive for the target in the high DNA background. 86 Organisms and cell lines may be tested as isolated DNA, or as pools of isolated DNA. Isolated DNA may 87 be combined into pools of up to 10 panel organisms, with each panel organism represented at 10 times 88 the AMDL, where possible. The combined DNA pools are tested in the presence (at 2 times the AMDL) 89 and absence of the target gene or gene fragment. If an unexpected result occurs, each of the individual 90 environmental organisms from a failed pool must be individually re-tested at 10 times the AMDL with and 91 without the target gene or gene fragment at 2x the AMDL in the candidate method DNA elution buffer. 92 93 DNA from organisms on this list that already appears in the inclusivity or exclusivity panel does not need 94 to be tested again as part of the environmental factors panel. Environmental Organisms

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Group 1

Aedes aegypti (ATCC /CCL-125(tm) mosquito cell line) Aedes albopictus (Mosquito C6/36 cell line) Dermatophagoides pteronyssinus (Dust mite -commercial source)

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Xenopsylla cheopis Flea (Rocky Mountain labs)

Drosophilia cell line

Musca domestica (housefly) ARS, USDA, Fargo, ND Gypsy moth cell lines LED652Y cell line (baculovirus)– Invitrogen

Cockroach (commercial source) 106 Tick (Amblyomma and Dermacentor tick species for F. tularensis detection assays) 1 107 Mus musculus (ATCC/HB-123) mouse 108 Rattus norvegicus (ATCC/CRL-1896) rat 109 Homo sapiens (HeLa cell line ATCC/CCL-2) human

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