Utilisation of Actiphage in combination with IS900 qPCR as a diagnostic tool for rapid determination of paratuberculosis infection status in small ruminant herds

Examination of fresh environmental samples for the presence of Mycobacterium avium subsp_ paratuberculosis (MAP) using an Actiphage–IS900 qPCR assay, direct IS900 qPCR, and a conventional phage amplification assay (PA)

Sample no.	Sample characteristics	Direct IS900 qPCR (copy/g)	Conventional PA (PFU/g)*	Actiphage–IS900 qPCR (copy/g)
1	Soil	5.42 × 102	4.80 × 102	–
2	Soil	1.09 × 102	2.18 × 102	–
3	Soil	1.02 × 102	2.88 × 102	–
4	Soil	4.20 × 102	–	–
5	Cobweb	1.20 × 102	–	–
6	Soil/plants	1.64 × 100	7.54 × 103	1.84 × 100
7	Soil	5.88 × 100	3.06 × 102	–
8	Soil	6.56 × 100	1.09 × 102	–
9	Soil	2.40 × 100	4.36 × 101	–

Examination of frozen environmental samples for the presence of Mycobacterium avium subsp_ paratuberculosis (MAP) using an Actiphage–IS900 qPCR assay, direct IS900 qPCR, a conventional phage amplification assay (PA) and culture

Sample no.	Sample characteristics	Time frozen	Direct IS900 qPCR (copy/g)*	Conventional PA (PFU/g)†	Culture*	Actiphage–IS900 qPCR
1	Soil	11 years	6.43 × 103	6.98 × 102	–	–
2	Soil	11 years	1.18 × 103	8.72 × 102	–	–
3	Soil	11 years	1.16 × 102	2.88 × 103	–	–
4	Soil	11 years	2.11 × 102	1.92 × 103	–	–
5	Soil	11 years	1.30 × 102	2.01 × 102	–	–
6	Soil	11 years	7.94 × 102	2.62 × 102	–	–
7	Soil	11 years	3.48 × 102	7.85 × 102	–	–
8	Roots	11 years	1.07 × 102	3.49 × 102	–	–
9	Soil	11 years	7.39 × 102	2.62 × 102	–	–
10	Roots	11 years	1.68 × 102	3.49 × 102	–	–
11	Soil	11 years	1.61 × 102	2.62 × 102	–	–
12	Roots	11 years	4.36 × 102	6.98 × 102	–	–
13	Roots	11 years	6.41 × 102	2.27 × 102	–	–
14	Soil	11 years	9.81 × 102	6.10 × 102	–	–
15	Soil	11 years	4.09 × 102	1.74 × 102	–	–
16	Roots	11 years	1.57 × 102	5.58 × 102	–	–
17	Soil	8 years	4.40 × 102	4.36 × 102	–	–
18	Detritus	8 years	1.25 × 102	4.36 × 101	–	–
19	Feed residues	8 years	1.94 × 102	–	–	–
20	Soil	11 years	2.67 × 101	8.72 × 101	–	–
21	Soil	11 years	7.26 × 101	1.13 × 103	–	–
22	Soil	11 years	8.93 × 101	5.67 × 102	–	–
23	Soil	11 years	6.11 × 101	3.49 × 103	–	–
24	Soil	11 years	7.95 × 101	1.74 × 102	–	–
25	Roots	11 years	6.44 × 101	5.23 × 101	–	–
26	Soil	11 years	7.90 × 101	8.72 × 100	–	–
27	Soil	11 years	2.70 × 101	1.40 × 102	–	–
28	Roots	11 years	9.91 × 101	1.74 × 102	–	–