Multiplexed-tandem PCR for the specific diagnosis of gastrointestinal nematode infections in sheep: an European validation study

Roeber, Florian; Morrison, Alison; Casaert, Stijn; Smith, Lee; Claerebout, Edwin; Skuce, Philip

doi:10.1186/s13071-017-2165-x

Table 1 Results of samples collected at Moredun, Scotland and Ghent, Belgium and tested by three different methods; multiplexed-tandem (MT)-PCR eggs (MT-PCR_(e)), MT-PCR larvae (MT-PCR_(l)) and larval culture (LC)

From: Multiplexed-tandem PCR for the specific diagnosis of gastrointestinal nematode infections in sheep: an European validation study

			MT-PCR gene copy number (calculated percentage)
Sample id (location)	EPG	Test	Tel	Haem	Trich	Cho	Oeso	Coc
M1	585	Mt-PCR_(e) ^a	693 (21)	0 (0)	2,572 (79)*	0 (0)	0 (0)	0 (0)
(Moredun)		MT-PCR_(l) ^a	0 (0)	0 (0)	120 (100)*	0 (0)	0 (0)	0 (0)
		LC %	83*	0	17	0		0
M2	90	Mt-PCR_(e) ^a	458 (94)	0 (0)	0 (0)	0 (0)	31 (6)	0 (0)
(Moredun)		MT-PCR_(l) ^a	2,062 (53)	0 (0)	45 (1)	1,788 (45)	44 (1)	0 (0)
		LC %	85	0	0	15		0
M3	243	Mt-PCR_(e) ^a	165 (29)	0 (0)	379 (66)	0 (0)	29 (5)	0 (0)
(Moredun)		MT-PCR_(l) ^a	261 (18)	0 (0)	1,123 (79)	0 (0)	43 (3)	0 (0)
		LC %	38	0	58	4		0
M4	30	Mt-PCR_(e) ^a	24 (3)	0 (0)	49 (6)	0 (0)	772 (91)	0 (0)
(Moredun)		MT-PCR_(l) ^a	0 (0)	0 (0)	295 (14)	0 (0)	1,799 (86)	0 (0)
		LC %	6	0	12	82		0
M5	176	Mt-PCR_(e) ^a	1,997 (86)*	0 (0)	26 (1)	309 (13)	0 (0)	0 (0)
(Moredun)		MT-PCR_(l) ^a	0 (0)	0 (0)	33 (1)	2,503 (99)*	0 (0)	0 (0)
		LC %	7	0	0	93*		0
M6	59	Mt-PCR_(e) ^b	0 (0)	0 (0)	146 (11)	1,241 (89)	0 (0)	0 (0)
(Moredun)		MT-PCR_(l) ^a	77 (0.3)	0 (0)	325 (1)	22,566 (99)	0 (0)	0 (0)
		LC %	4	0	1	94		0
M7	13	Mt-PCR_(e) ^b	113 (100)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
(Moredun)		MT-PCR_(l) ^a	631 (91)	0 (0)	61 (9)	0 (0)	0 (0)	0 (0)
		LC %	100	0	0	0		0
M8	11	Mt-PCR _(e)	16,693 (100)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
(Moredun)		MT-PCR_(l) ^b	514 (76)	0 (0)	0	165 (24)	0 (0)	0 (0)
		LC %	97	0	0	3		0
M9	171	Mt-PCR_(e) ^b	3,994 (82)	0 (0)	602 (12)	133 (3)	158 (3)	0 (0)
(Moredun)		MT-PCR_(l) ^a	1,075 (79)	0 (0)	245 (18)	0 (0)	42 (3)	0 (0)
		LC %	79	0	15	6		0
M10	36	Mt-PCR _(e)	33,346 (30)	0 (0)	47,858 (43)**	24,912 (22)	2,709 (3)	2,100 (2)
(Moredun)		MT-PCR_(l)	19 (0.03)	0 (0)	8,189 (14)	42,319 (72)*	0 (0)	8,100 (14)
		LC %	34	0	2	14		50*
M11	720	Mt-PCR _(e) ^b	358 (1)	27,983 (92)	130 (0.43)	1,792 (6)	253 (1)	0 (0)
(Moredun)		MT-PCR_(l) ^b	0 (0)	4,309 (99)	62 (1)	0 (0)	0 (0)	0 (0)
		LC %	0 (0)	98	0	1		1
B1	176	Mt-PCR_(e)	10,762 (85)	15 (0.12)	0 (0)	0 (0)	0 (0)	1,900 (15)
(Ghent)		MT-PCR_(l)	14,617 (78)	40 (0.21)	0 (0)	0 (0)	0 (0)	4,000 (21)
		LC %	92	0	8	0	0	0
B2	2	Mt-PCR_(e)	13,157 (8)	135,922 (84)	1,793 (1)	11,398 (7)	0 (0)	0 (0)
(Ghent)		MT-PCR_(l)	1,619 (1)	79,775 (67)	887 (1)	36,547 (31)	0 (0)	0 (0)
		LC %	10	68	5	17		0
B3	176	Mt-PCR_(e)	8,262 (12)	34,497 (51)*	25,297 (37)	0 (0)	0 (0)	0 (0)
(Ghent)		MT-PCR_(l)	3,887 (84)*	0 (0)	752 (16)	0 (0)	0 (0)	0 (0)
		LC %	99*	1	0	0		0
B4	72	Mt-PCR_(e)	6,914 (71)*	406 (4)	16 (0.16)	2,436 (25)	0 (0)	0 (0)
(Ghent)		MT-PCR_(l)	4,154 (13)	21,880 (68)*	2,275 (7)	0 (0)	0 (0)	3,900 (12)
		LC %	17	83*	0	0		0
B5	200	Mt-PCR_(e)	22,039 (100)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
(Ghent)		MT-PCR_(l)	172,336 (100)	0 (0)	0 (0)	0 (0)	0 (0)	0 (0)
		LC %	68	0	32	0		0
B6	986	Mt-PCR_(e)	6,741 (5)	39,499 (30)	220 (0.17)	83,044 (63)*	0 (0)	3,100 (2)
(Ghent)		MT-PCR_(l)	175 (26)	494 (74)*	0 (0)	0 (0)	0 (0)	0 (0)
		LC %	17	74*	6	0		3
B7	1584	Mt-PCR_(e)	153,756 (20)	531,837 (70)	0 (0)	72,565 (10)	945 (0.12)	0 (0)
(Ghent)		MT-PCR_(l)	1,195,981 (10)	9,266,891 (77)	1,955 (0.02)	1,436,634 (12)	83,435 (1)	5,900 (0.05)
		LC %	30	65	0	5		0
B8	804	Mt-PCR_(e)	267,862 (11)	39 (0.00)	566,001 (24)	0 (0)	1,414,909 (59)*	156,200 (6)
(Ghent)		MT-PCR_(l)	8,309 (3)	56 (0.02)	84,321 (28)	0 (0)	28,087 (9)	180,300 (60)*
		LC %	24	1	4	10		61*

^aSamples tested at 1/50 dilution
^bSamples tested at 1/100 dilution
*Samples with discrepant results
For every sample, the microscopy results are given as eggs per gram of faeces (EPG). The species/genera-specific results of the MT-PCR for T. circumcincta (Tel), Haemonchus spp. (Haem), Trichostrongylus spp. (Trich), C. ovina (Cho), Oesophagostomum spp. (Oeso) and Co. curticei (Coc) are shown as gene copy number or as calculated percentage (%). Discrepancies between the results of the three diagnostic methods are indicated by an asterisk

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