September 2015 SPADA Meeting

Reference Materials

• Any material being used to qualify the “assay(s)” •

Filters or liquid from liquid samplers (if not standardized) • Soils used to challenge prospective “assay(s)” • Other matrices identified as a matrix for which the assay will be validated • Cultures used to challenge the assay(s)

• Certified or Noncertified Reference Materials • What should be a reference material •

Key organisms in the inclusivity panel and exclusivity panel • Matrices of interest (air samples or air sample equivalents)

• SuggestedReference Material for Air sample testing‐ • Dust from air conditioners, air conditioner chases, filters from commercial and residential buildings in multiple locations. Dust should be tested by existing test methods to determine  cross reactivity for any B. anthracis  targets • Accepted dust was sieved to get multiple samples of dust from distinct origins • Each was tested by culture and PCR currently being used for BA detection (3 distinct reactions) • Equalvolumes of each dust were homogenized for 72 hours in a rolling drum • Homogenized dust was tested using PCR and culture for BA • Homogenized dust was placed in 100 ml serum bottles, frozen and lyophilized • The dust was characterized for particle size and background organisms  • This SMPR had • 15B.anthracis reference cultures (1certified) • 20otherBacillus cultures (1certified) • 87environmental biological sampleDNAs (0certified) • 25soils (0certified) • 22powdersand chemicals (0certified)

Bacillus anthracis Inclusivity Strains

Number

Strain

Characteristics

BA1

Canadian bison pXO1+ , pXO2+, VNTR genotype group A1a

BA2

pXO1+ , pXO2‐,  VNTR genotype group A1b

V770‐NP‐1R

• The study included DNA from 15  B. anthracis strains tested at the  AMDL (2000 genome  equivalents/mL or 11 pg/mL) or  lower and DNA from 20 near  i hb i t t d t 10 ne g or organ sms es e a   times the AMDL, or 110 pg/mL

BA3

pXO1+ , pXO2+, VNTR genotype group A2

PAK‐1

BA4

pXO1+ , pXO2+, VNTR genotype group A3a

BA1015

BA5

pXO1+ , pXO2+, VNTR genotype group A3b

Ames

BA6

pXO1+,  pXO2+, VNTR genotype group A3c

K3

BA7

pXO1+,  pXO2+, VNTR genotype group A3d

Ohio ACB

SK‐102  (Pakistan) Vollum 1B

BA8

pXO1+ , pXO2+, VNTR genotype group A4

BA9

pXO1+ , pXO2+, VNTR genotype group A4

BA10

pXO1+ , pXO2+, VNTR genotype group B1

BA1035

BA11

pXO1+, pXO2+, VNTR genotype group B2

RA3

2002013094  (240)

BA12

pXO1+, pXO2+, VNTR genotype group C

BA13

pXO1‐ , pXO2+; VNTR genotype group A1a

Pasteur

BA14

pXO1+ , pXO2‐, VNTR genotype group A3b

Sterne

BA15

Turkey #32 pXO1+, pXO2+, VNTR genotype group A1b