MT-CO1 and infection: Finally, the analytical sensitivity was confirmed for both the cox1-triplex and the ftsZ duplex qPCRs in detecting the infection by D. immitis (Additional file 9: Table S6).The detection limit was up to 4.03 × 10−2 and 4.03 × 10−1 mf/ml, respectively (corresponding to 2.01 × 10−3 and 2.01 × 10−2, respectively) mf/5 μl, qPCR efficiency was 104.8 and 100.5%, respectively, slopes were − 3.212 and − 3.309, respectively.