AOAC SPADA Meeting

PSET Results (Ebola Assays)

Assay #

True-negative Report*

Assay ID

AssayHit

GeneTarget

Percentage of perfect True-positive

False-positive

Percentage of perfect Amplicon hits

False-negative

PerfectAssayHits

Species/Strain of hits

Perefect ampliconhit

Amplicon length (bps)

IntendedTarget Species

1 2 3 4 4 5 6 7

Sig1

EBOV EBOV EBOV EBOV EBOV EBOV EBOV EBOV EBOV EBOV EBOV EBOV

NP NP NP NP NP NP NP NP GP GP GP GP NP NP GP GP NP GP GP GP NP NP NP NP NP GP L L L L

EBOV 124 135 132 97.8 135 11

8.1

135 0

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0

0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 9 0 0 0 8 0 0 0

Pass Pass Pass Pass Pass Pass Pass Fail Pass Fail Pass Pass Pass Pass Pass Pass Pass Pass Pass Fail Pass

EbolaZaire-MGB

EBOV EBOV EBOV SUDV

76 135 134 99.3 135 124 91.9 135 0 49 135 135 100.0 135 125 92.6 135 0

Sig3

EboZNP EboZNP ZAI-NP

80 136 22

16.2 136 19

14.0 136 0

80

0

0

0

11

0

0

0

0

11

EBOV 268 148 123 83.1 148 10

6.8 7.4

148 0 148 0

EbolaMGB-EBOV

EBOV EBOV

79 148 23 70 148 32

15.5 148 11 21.6 148 32

ENZ

21.6 148 0

• Most of the assays passed with the relaxed criteria (90/90; 85/90 rule) (3 failed- false positive or false negative) • Only 3 passed 100/100 rule

10 10 12 13 16 16 16 17 20 21 22 23

EBO-GP-1 EBO-GP-1 ZebovGP

EBOV 579 152 RESTV 579 12

0 0

0 0

152

5 0

3.3

152 0

0

0

0

0

12

EBOV EBOV

64 153 13 80 144 13

8.5 9.0

153 13 153 13 135 12

8.5 8.5 8.9

153 0 144 9 135 0

EbolaZaire-TM

FiloAB FiloAB FiloAB GAB-1

pan-Filo pan-Filo pan-Filo EBOV BDBV BDBV TAFV TAFV RESTV

EBOV 419 135 MARV 419 55 SUDV 419 12

0 0 0 1 1 1 2 8 2 0 2

0 0 0

0 0

0 0

0 0

55 12

0 0

EBOV 353 135 17

12.6 135 12

8.9

135 0

19 EbolaBDBV-MGB

BDBV BDBV TAFV TAFV

74 74 64 79

5 5 2 2 8 8 0 8

20.0 20.0 50.0

5 5 2 2 8 8 9 8

1 1 1 2 3 2 0 2

20.0 20.0 50.0

5 5 2 2 8 8 0 8

0 0 0 0 0 0 0 0 1 0 0 0 0 0 0

EbolaBDBV-TM EbolaTAFV-MGB EbolaTAFV-TM

100.0 100.0

100.0

Reston

RESTV 337 RESTV 97

37.5 25.0

24 Ebola-MGB-RESTV RESTV 24 Ebola-MGB-RESTV RESTV

25.0

EBOV

97

0

0

25

EbolaReston-TM

RESTV VP40 RESTV 80

25.0 90.9 90.9

25.0 83.3 90.9

26 EbolaReston-MGB RESTV

RESTV 55

11 11

10 10

12 11

10 10

11 11

27 27 27

EbolaSudan-MGB EbolaSudan-MGB EbolaSudan-MGB

SUDV SUDV SUDV SUDV SUDV SUDV

SUDV

80

RESTV 80

0 0

0 0 7

0 0

8

0 0 2

0 0

0 0

EBOV SUDV SUDV SUDV

80 81 89 77

148

0 148

28 EbolaMGB-SUDV

11 11 14

63.6 90.9 64.3

11 11 14

18.2 90.9 64.3

11 11 14

Pass Pass Pass

29 30

Sudan

10

10

EbolaSudan-TM

9

9

2018_10_17_SPADA_Brief

UNCLASSIFIED

19

19

Heat map of assay hits to Gen Bank entries • Most of the EBOV assays have less than perfect matches to many GenBank entries • Species specific assays are mostly specific with relaxed criteria • Cross reactivity is seen with pan assays and specific assays • Assay positive but amplicon negative hits are due to extensive variation and pan assays • Amplicon positive but assay negative hits are usually genetic drift

2018_10_17_SPADA_Brief

UNCLASSIFIED

20

20