A one-step multiplex qPCR assay for simultaneous identification and quantification of Leishmania martiniquensis and Leishmania orientalis/Leishmania chancei and detection and quantification of trypanosomatids in clinical samples

Open Access

Table 2

Analytical specificity of the multiplex qPCR assay.

DNA samples	Quantification cycle (Cq) (Mean ± SD)
DNA samples	ITS1-L. mar-HEX	ITS1-L. ori/cha-FAM	ITS1-Tryps- Texas Red	RP-human-Cy5
L. martiniquensis (LV760) + human DNA^a	12.77 ± 0.12	ND^d	11.36 ± 0.14	25.58 ± 0.12
L. martiniquensis (LSCM1) + human DNA^a	12.77 ± 0.05	ND	12.47 ± 0.08	25.45 ± 0.09
L. martiniquensis (LSCM1-6) + human DNA^a	11.11 ± 0.11	ND	14.20 ± 0.04	25.85 ± 0.06
L. martiniquensis (AmpBRP2i) + human DNA^a	11.12 ± 0.15	ND	13.12 ± 0.08	25.63 ± 0.19
L. martiniquensis (LSCM2) + human DNA^a	13.49 ± 0.02	ND	13.53 ± 0.11	25.89 ± 0.15
L. orientalis (LSCM4) + human DNA^a	ND	11.31 ± 0.14	13.78 ± 0.01	25.52 ± 0.02
L. orientalis (PCM2) + human DNA^a	ND	16.32 ± 0.21	11.12 ± 0.04	25.18 ± 0.12
L. chancei + human DNA^a	ND	16.44 ± 0.06	17.63 ± 0.04	25.65 ± 0.20
L. aethiopica + human DNA^a	ND	ND	10.64 ± 0.12	25.39 ± 0.10
L. amazonesis + human DNA^a	ND	ND	13.24 ± 0.21	25.30 ± 0.06
L. donovani + human DNA^a	ND	ND	16.78 ± 0.03	25.74 ± 0.08
L. infantum + human DNA^a	ND	ND	15.28 ± 0.01	25.24 ± 0.10
L. major + human DNA^a	ND	ND	14.46 ± 0.21	25.40 ± 0.04
L. mexicana + human DNA^a	ND	ND	12.71 ± 0.16	25.19 ± 0.08
L. tropica + human DNA^a	ND	ND	13.51 ± 0.16	25.18 ± 0.14
L. braziliensis + human DNA^a	ND	ND	15.06 ± 0.02	25.87 ± 0.02
L. guyanesis + human DNA^a	ND	ND	14.13 ± 0.05	25.34 ± 0.18
L. panamensis + human DNA^a	ND	ND	15.78 ± 0.14	25.22 ± 0.11
T. brucei + human DNA^a	ND	ND	25.31 ± 0.05	24.49 ± 0.05
Crithidia sp. (CLA-KP1) + human DNA^a	ND	ND	13.27 ± 0.19	25.13 ± 0.21
Crithidia sp. (CLA-KP4) + human DNA^a	ND	ND	15.27 ± 0.15	25.42 ± 0.11
Crithidia sp. (CLA-TR2) + human DNA^a	ND	ND	14.11 ± 0.12	25.39 ± 0.14
Crithidia sp. (CLA-TR3) + human DNA^a	ND	ND	14.74 ± 0.22	25.40 ± 0.01
P. vivax^b	ND	ND	ND	25.30 ± 0.16
P. falciparum ^b	ND	ND	ND	25.45 ± 0.11
M. tuberculosis^c	ND	ND	ND	ND

^aArtificial mixture; ^bClinical sample; ^cCulture sample; ^dND = not detected.

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