Development and validation of a long-read metabarcoding platform for the detection of filarial worm pathogens of animals and humans

Table 4 Modified Knott’s test versus metabarcoding assay agreement statistics for the two most common filarial worm pathogens identified from 100 Sri Lanka dog blood samples

Filarial worm	Modified Knott’s test	Metabarcoding assay		Total agreement (%)	Kappa (95% CI)	Agreement	Kappa SE
Filarial worm	Modified Knott’s test	NEG	POS	Total agreement (%)	Kappa (95% CI)	Agreement	Kappa SE
Brugia spp.	NEG	81	17	83	0.16 (0.059–0.261)	Poor	0.003
Brugia spp.	POS	0	2	83	0.16 (0.059–0.261)	Poor	0.003
Dirofilaria spp.	NEG	47	34	65	0.317 (0.246–0.388)	Fair	< 0.001
Dirofilaria spp.	POS	1	18	65	0.317 (0.246–0.388)	Fair	< 0.001

POS = positive, NEG = negative, CI = confidence intervals, SE = standard error. Agreement level defined as poor if coefficient (k) is ≤ 0.20, fair agreement if 0.21 ≤ k ≤ 0.40, moderate agreement if 0.41 ≤ k ≤ 0.60, substantial agreement if 0.61 ≤ k ≤ 0.80, and high agreement if k > 0.81. Genus-level taxonomic classifications are shown as the modified Knott’s test can only identify microfilaria to this taxonomic level, whilst our metabarcoding protocol could classify down to species level in all instances

ISSN: 1471-2180