Multiplex-RT-PCR-ELISA panel for detecting mosquito-borne pathogens: Plasmodium sp. preserved and eluted from dried blood spots on sample cards

Koliopoulos, Philip; Kayange, Neema Mathias; Daniel, Tim; Huth, Florian; Gröndahl, Britta; Medina-Montaño, Grey Carolina; Pretsch, Leah; Klüber, Julia; Schmidt, Christian; Züchner, Antke; Ulbert, Sebastian; Mshana, Steven E.; Addo, Marylyn; Gehring, Stephan

doi:10.1186/s12936-021-03595-4

Table 2 Multiplex-RT-PCR-ELISA: primers and probes

From: Multiplex-RT-PCR-ELISA panel for detecting mosquito-borne pathogens: Plasmodium sp. preserved and eluted from dried blood spots on sample cards

	Family	No	Pathogens and prevalence	Primer	5′-3′ sequence	Concentration ( pm/µl)	Literature^a
Arboviruses	Flaviviridae	1	Dengue virus (DENV) [8, 42, 61, 62]	C-prm D1	TCAATATGCTGAAACGCGAGAGAAACCG	5.5	[25]^b,c
			Dengue virus (DENV) [8, 42, 61, 62]	D2	TTGCACCAACAGTCAATGTCTTCAGGTTC	5.5
			DENV-1^d	rTS1 (DEN-1)	CCCGTAACACTTTGATCGCT	5.5
			DENV-2^d	mTS2 (DEN-2)	CGCCACAAGGGCCATGAACAGTTT	5.5
			DENV-3^d	TS3 (DEN-3)	TAACATCATCATGAGACAGAGC	5.5
			DENV-4^d	rTS4 (DEN-4)	TTCTCCCGTTCAGGATGTTC	5.5
		2	West Nile virus (WNV) [63,64,65]	WNV lancio F	CAGACCACGCTACGGCG	5.5	[66]^b
				WNV lancio R	CTAGGGCCGCGTGGG	5.5
				WNV lancio P	TCTGCGGAGAGTGCAGTCTGCGAT	10
		3	Zika virus (ZIKV) [67, 68]	ZVforw	CAGCTGGCATCATGAAGAAYC	5.5	[70]^b
				ZVrev1	CACTTGTCCCATCTTCTTCTCC	5.5
				ZVrev2	CACCTGTCCCATCTTTTTCTCC	5.5
				ZVprobe	CYGTTGTGGATGGAATAGTGG	10
		4	Yellow Fever virus (YFV) [69,70,71]	YFV-F	GCACGGATGTAACAGACTGAAGA	5.5	[72]^b
				YFV-R	CCAGGCCGAACCTGTCAT	5.5
				YFV-probe	CGACTGTGTGGTCCGGCCCTC	10
	Togaviridae (Alphaviruses) [37]	5	Semliki Forest virus (SFV) [64]	SFV-F	ACAGACTGTCACTGAGCAG	5.5	[75]^b
				SFV-R	AGCTCCACGTCATCATTGAG	5.5
				SFV-probe	GTGACCATCTACTGCAGAGA	10
		6	O´nyong-nyong virus (ONNV) [74,75,76]	ONNV-F	GCAGGGAGGCCAGGACAGT	5.5	[79]^b
				ONNV-R	GCCCCTTTTTCYTTGAGCCAGTA	5.5
				ONNV-probe	TGTATTGCTCCTGCCGCTGG	10
		7	Chikungunya virus (CHIKV) [42, 76, 78, 79]	ChikS	TGATCCCGACTCAACCATCCT	5.5	[82]^b
				ChikAs	GGCAAACGCAGTGGTACTTCCT	5.5
				ChikP	TCCGACATCATCCTCCTTGCTGGC	10
	Bunyaviridae	8	Rift Valley fever virus (RVFV) [81, 82]	RVF-F	TGAAAATTCCTGAGACACATGG	5.5	[79]^b
				RVF-R	ACTTCCTTGCATCATCTGATG	5.5
				RVF-probe	CACAAGTCCACACAGGCCCCTTACAT	10
Protozoa	Malaria (Plasmodium sp.)	9	Plasmodium sp. (MAL) [83]	Plasmo Plu3 F	GCTCTTTCTTGATTTCTTGGATG	5.5	[26]^b
				Plasmo Plu3 R	AGCAGGTTAAGATCTCGTTCG	5.5
				Plasmo Plu3 P	ATGGCCGTTTTTAGTTCGTG	10
			P. falciparum [84]	FAL-F	CTTTTGAGAGGTTTTGTTACTTTGAGTAA	5.5	[51]^e
				FAL-R	TATTCCATGCTGTAGTATTCAAACACAA	5.5
				FAL-probe	TGTTCATAACAGACGGGTAGTCATGATTGAGTTCA	10
			P. vivax [85]	VIV-F	ACGCTTCTAGCTTAATCCACATAACT	5.5	[88]^e
				VIV-R	ATTTACTCAAAGTAACAAGGACTTCCAAGC	5.5
				VIVAX-probe	TTCGTATCGACTTTGTGCGCATTTTGC	10
			P. malariae [87, 88]	Pm-1	AGTTAAGGGAGTGAAGACGATCAGA	5.5	[89]^e
				Pm-2	CAACCCAAAGACTTTGATTTCTCATAA	5.5
				Pm-probe	ATGAGTGTTTCTTTTAGATAGC	10

^aThe primers and probes listed were obtained from the literature cited
^bIncluded in the multiplex-RT-PCR-ELISA
^cUsed to type DENV1-4 according to the method of Chien et al. [25]
^dProbes for DENV1-4 consisted of the respective biotinylated primers
^eUsed to differentiate Plasmodium sp. by multiplex-RT-PCR-ELISA

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