Table 1.

Cross-reactivity results of the multiplexed Taqman PCR assays for detecting variola and other orthopoxviruses

Species/sample	Isolate	Conc. [Fg]	Dry Ct FAM	Dry Ct TET	Liquid Ct FAM	Liquid Ct TET
Orthopoxvirus samples
Camelpox virus	Somalia	1000	0/2	2/2	0/2	2/2
Cowpox virus	Brighton	1000	0/2	2/2	0/2	2/2
J7R Cloned DNA	BHS	100	5/5	5/5	5/5	5/5
J7R Cloned DNA	BHS	10	8/8	8/8	8/8	8/8
J7R Cloned DNA	BHS	0.1	11/12a	12/12	12/12	11/12a
Monkeypx virus	Zaire 96(I-16)	1000	0/4	0/4	0/4	0/4
Monkeypx virus	Zaire 96(I-16)	100	0/5	0/5	0/5	0/5
Monkeypx virus	Zaire 96(I-16)	10	0/6	5/6a	0/6	5/6a
Myxoma virus	CDC	1000	0/2	0/2	0/2	0/2
Racconpox virus	CDC	1000	0/2	2/2	0/2	2/2
Rabbitpox virus	CDC	1000	0/2	2/2	0/2	2/2
Skunkpox virus	CDC	1000	0/2	0/2	0/2	0/2
Tanapox virus	CDC	1000	0/2	2/2	0/2	2/2
Vaccinia virus	CPN	1000	0/2	2/2	0/2	2/2
Bacterial samples
Bacillus anthracis	4728	1000	0/2	0/2	0/2	0/2
Francisella tularensis	NA	1000	0/2	0/2	0/2	0/2
Yersinia pseudotuberculosis	690	1000	0/2	0/2	0/2	0/2
Stpahylococcus hominis	27844	1000	0/2	0/2	0/2	0/2
Neisseria lactamica	23970	1000	0/2	0/2	0/2	0/2
Haemophlius influenzae	10211	1000	0/2	0/2	0/2	0/2
Staphylococcus aureus	25923	1000	0/2	0/2	0/2	0/2
Listeia monocytogenes	15313	1000	0/2	0/2	0/2	0/2
Streptococcus pyogenes	19615	1000	0/2	0/2	0/2	0/2
Clostridium perfringens	13124	1000	0/2	0/2	0/2	0/2
Viral samples
Black Creek Canal virus	39179	100	0/2	0/2	0/2	0/2
Ebola Sudan virus	Boniface	100	0/2	0/2	0/2	0/2
Ebola Zaire virus	Zaire95	100	0/2	0/2	0/2	0/2
Hantaan virus	76-118	100	1/2b	0/2	0/2	0/2
Lassa virus	Josiah	100	0/2	0/2	0/2	0/2
Dengue 3	CH3489	100	0/2	0/2	0/2	0/2
Eastern Equine Encephalitis	FL-4679	100	0/2	0/2	0/2	0/2
Rift Valley fever virus	ZH 548	100	0/2	0/2	0/2	0/2
Venezuelan e. encephalitis	PE-4.0904	100	0/2	0/2	0/2	0/2
West Nile	Crow397-99	100	0/2	0/2	0/2	0/2
Yellow Fever	Assibe	100	0/2	0/2	0/2	0/2
Other samples
Human genome DNA	N/A	1000	0/2	0/2	0/2	0/2

Comparison of the performance of the assay with dried (Dry FAM and Dry TET) and liquid (Liquid FAM and Liquid TET) reagents showing the number of positive samples over the total number of samples tested. The sensitivity and specificity were determined as described in Section 2. The results indicate that the sensitivity and specificity of the assay with dried PCR reagents, were 96% and 98%, respectively, and the sensitivity and specificity of the liquid reagents were 100%.

^aOne sample has a C_t value below the threshold.

^bFalse positive.