SPADA Working Groups ~ April Meeting Book

Draft, Do Not Distribute

Table IV: Environmental Factors Panel For Validating PCR Detectors For Biothreat 79 Agents 80 81 [Adapted from the Environmental Factors Panel approved by SPADA on June 10, 2010.] 82 83 The Environmental Factors Panel is intended to supplement the biothreat agent near- neighbor 84 exclusivity testing panel, and it should be applicable to all PCR biothreat agent detection assays. 85 The panel criteria are divided into two main groups – the matrix panel of unknown 86 environmental samples (Part 1); and the environmental panel of identified environmental 87 organisms (Part 2). This panel will test for potential cross-reactive amplification and/or PCR 88 inhibitors. 89

90

Part 1:

91 92 Environmental Matrix Samples - Aerosol Environmental Matrices

93 94 95 96 97 98 99

o The aerosol environmental matrix pools should be used to confirm that there is no detection with the method used i.e. there is no cross reactivity of the target assay

with unknown environmental organisms.

o The aerosol environmental matrix pools should also be tested with the target fragment at the AMDL to confirm the filter pool does not interfere with detection by 100 101 · Method developers should obtain environmental matrix samples that are representative 102 and consistent with the collection method that is anticipated to be utilized in generating the 103 sample being analyzed. This includes considerations that may be encountered when the 104 collection system is deployed operationally such as collection medium, duration of 105 collection, diversity of geographical areas that will be sampled, climatic/environmental 106 conditions that may be encountered and seasonal changes in the regions of deployment. 107 Justifications for the selected conditions that were used to generate the environmental 108 matrix and limitations of the validation based on those criteria must be documented. the method used. o Method developers will test the environmental matrix for interference with sufficient samples to achieve 95% probability of detection. o Cross-reactivity testing will include sufficient samples and replicates to ensure each environmental condition is adequately represented .

109 110 111 112 113 114 115 116

6 Draft SMPR Coxiella burnetii V3