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 1

Primers and probes for the ITS1 and human RNase P targets used in the developed multiplex qPCR assay.

Target	Name of primer-probe set	Sequence	Tm (°C)	Fluorescent Dye/Amplicon size (bp)
ITS1	ITS1-L. mar^a	Forward: CCACATACACAAACACAGCAATA (Sense)	62	HEX/149
		Probe: GCCAAATGCCGCGCGTATACAG (Sense)	68
		Reverse: GAGAGAAAGAGCCGTAACGAA (AntiSense)	62
	ITS1-L. ori/cha^b	Forward: GGGAGGTGTMTCTCTCTTT (Sense)	60	FAM/146
		Probe: AGATARCGCCTTTCCCACATACACA (Sense)	68
		Reverse: TACGCYCGGTGTTTATATG (AntiSense)	60
	ITS1-Tryps^c	Forward: CGGTGTGTTGTGGATAACGG (Sense)	63	Texas Red/77
		Probe: TAACGTGTCGCGATGGATGACTTGG (Sense)	68
		Reverse: CTGCGTTCTTCAACGAAATAGGA (AntiSense)	63
Human RNase P	RP-human^d	Forward: TCAGCATGGCGGTGTTT (Sense)	62	Cy5/81
		Probe: TTCTGACCTGAAGGCTCTGCGC (Sense)	68
		Reverse: CGGCTGTCTCCACAAGTC (AntiSense)	62

^aFor detection of L. martiniquensis; ^bFor detection of L. orientalis/L. chancei; ^cFor detection of trypanosomatids; ^dInternal control

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