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

Table 3 Conventional PCR with Sanger sequencing versus metabarcoding assay agreement statistics for the two most common filarial worm pathogens identified from 100 Sri Lanka dog blood samples

Filarial worm	cPCR and Sanger sequencing	Metabarcoding assay		Total agreement (%)	Kappa (95% CI)	Agreement	Kappa SE
Filarial worm	cPCR and Sanger sequencing	NEG	POS	Total agreement (%)	Kappa (95% CI)	Agreement	Kappa SE
Brugia sp. SL genotype	NEG	81	11	89	0.541 (0.425–0.657)	Moderate	< 0.001
Brugia sp. SL genotype	POS	0	8	89	0.541 (0.425–0.657)	Moderate	< 0.001
Dirofilaria sp. ‘hongkongensis’	NEG	47	21	78	0.566 (0.491–0.641)	Moderate	< 0.001
Dirofilaria sp. ‘hongkongensis’	POS	1	31	78	0.566 (0.491–0.641)	Moderate	< 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

ISSN: 1471-2180