Development of a Large-Scale Pathogen Screening Test for the Biosafety Evaluation of Canine Mesenchymal Stem Cells

Pekker, Emese; Priskin, Katalin; Szabó-Kriston, Éva; Csányi, Bernadett; Buzás-Bereczki, Orsolya; Adorján, Lili; Szukacsov, Valéria; Pintér, Lajos; Rusvai, Miklós; Cooper, Paul; Kiss-Tóth, Endre; Haracska, Lajos

doi:10.1186/s12575-023-00226-x

Table 5 Primers and probes used for extraneous agent detection

From: Development of a Large-Scale Pathogen Screening Test for the Biosafety Evaluation of Canine Mesenchymal Stem Cells

	Infectious agent	Detection method	Primer name	Primer sequence (5′-3′)	Product size (bp)	Reference
RNA viruses	CDV	Conventional PCR	CDV_N_F768	AACAGRRATTGCTGAGGACYTAT	290	[32]
	CDV	Conventional PCR	CDV_N_R1057	TCCARRRATAACCATGTAYGGTGC	290	[32]
	CPIV	Conventional PCR	CPIV_N_F428	GCCGTGGAGAGATCAATGCCTAT	186	[32]
	CPIV	Conventional PCR	CPIV_N_R614	GCGCAGTCATGCACTTGCAAGT	186	[32]
	CIV	Conventional PCR	CIV_M_F151	CATGGARTGGCTAAAGACAAGACC	126	[32]
	CIV	Conventional PCR	CIV_M_R276	AGGGCATTTTGGACAAAKCGTCTA	126	[32]
	Rabies virus	Conventional PCR	JW12-F	ATGTAACACCYCTACAATG	606/582	[33]
			JW6 UNI-R	ARTTVGCRCACATYTTRTG
			JW 10 UNI-R	GTCATYARWGTRTGRTGYTC
	CCoV	Conventional PCR	CoV_16053_F	GGTTGGGAYTAYCCTAARTGTGA	452	[32]
	CCoV	Conventional PCR	CoV_Pan_16510_R	TTATARCAVACAACNCCATCATCA	452	[32]
DNA viruses	CAV-1	Conventional PCR	CadV_E3_F25073	TATTCCAGACTCTTACCAAGAGG	545	[32]
	CAV-1	Conventional PCR	CadV_E3_R25623	ATAGACAAGGTAGTARTGYTCAG	545	[32]
	CPV	Conventional PCR	CPV-F	AAGACGTGCAAGCGAGTCC	337	[34]
	CPV	Conventional PCR	CPV-R	GAGCGAAGATAAGCAGCGTAA	337	[34]
	CaHV-1	Conventional PCR	CaHV_GBF439	ACAGAGTTGATTGATAGAAGAGGTATG	136	[32]
	CaHV-1	Conventional PCR	CaHV_GBR574	CTGGTGTATTAAACTTTGAAGGCTTTA	136	[32]
	SuHV1	Conventional PCR	gB-Taq-F	CTCCTGCCGCACCTGAAG	92	[35]
	SuHV1	Conventional PCR	gB-Taq-R	GTCTGGAAGCGGTAGAAGCC	92	[35]
Bacteria	Leptospira interrogans	Conventional PCR	LepN-1	CTGGCCTAAAACTGACGCTGA	171	[20]
	Leptospira interrogans	Conventional PCR	LepN-2	CTTTCACTCTTGCGAGCATAG	171	[20]
	Borrelia	Real-time PCR	BorP66F	GCAATTTTAGCATCTTTTGGAG	106	[36]
	Borrelia	Real-time PCR	BorP66R	GATCTATTCCAAAATCRGTWCC	106	[36]
	Brucella canis	Conventional PCR	B2N-1	GTCGCGGATTCTACCTCACCT	281	[20]
	Brucella canis	Conventional PCR	B2N-2	TAAGCAGGTAAGAGGCAATTT	281	[20]
	Ehrlichia spp.	Real-time PCR	Ehr16SF	GGCACGTAGGTGGACTA	101	[36]
	Ehrlichia spp.	Real-time PCR	Ehr16SR	TTCCGCTATCCTCTTTCGAC	101	[36]
	Anaplasma	Real-time PCR	Ana/Ehr16SF	ACACATGCAAGTCGAACG	145	[36]
	Anaplasma	Real-time PCR	Ana/Ehr16SR	CCCCCGCAGGGATTATACA	145	[36]
	Neorickettsia	Real-time PCR	groel-1500F	ATAGATCCAGCKAAGGTAGTGCGTGT	148	[37]
	Neorickettsia	Real-time PCR	groel-1620R	TTCCACCCATGCCACCACCAGGCATCATTG	148	[37]
	Rickettsia	Real-time PCR	Ric16SF	TCCTAGTGTAGAGGTGAAATTCTTA	178	[36]
	Rickettsia	Real-time PCR	Ric16SR	GAAACCGAAAGAGAATCTTCCGAT	178	[36]
	Bartonella	Real-time PCR	ssrA-F	GCTATGGTAATAAATGGACAATGAAATAA	298	[38]
	Bartonella	Real-time PCR	ssrA-R	GCTTCTGTTGCCAGGTG	298	[38]
	Mycoplasma	Real-time PCR	Myco16sQF1	GCAAAGCTATAGAGATATAGTAGAGGT	106/107 and 100 bp for internal control	[39]
			Myco16sQF2	GCRAAGCTATAGARATATAGTGGAGGT
			Myco16sQF3	GCAATGCTATAGAGATATAGCGGAGGT
			Myco16sQF4	GCAAAGCTATGGAGACATAGTGGAGGT
			Myco16sQF5	GCAAAGTTATGGAAACATAATGGAGGT
			Myco16sQF6	GCGACGCTATAGAAATATAGTTGAGGT
			Myco16sQF7	GCAAAGGCTTAGAAATAAGTTCGGAGGC
			Myco16sQF8	GCAAAGCTATAGAAATATAGTAGAGGT
			Myco16sQR	GTTGCGYTCGTTGCRGGAC
			Myco16sQProbe	FAM-TGGTGCATGGTTGTC-MGB
			Myco16sQICProbe	VIC-CACGCCGTAAACGA-MGB
Protozoa	Leishmania	Real-time PCR	LEISH-2	ACCCCCAGTTTCCCGCC	119	[40]
	Leishmania	Real-time PCR	LEISH-1	AACTTTTCTGGTCCTCCGGGTAG	119	[40]
	Babesia	Conventional PCR	B-BM	GAATCTAAACCCTTCCCAGAGTATC	381/408	[41]
			B-BDV	TGACCTAAACCCTCACCAGAGTAAC
			B-BDV2	TGACCCAAACCCTCACCAGAGTARC
			B-rev	GCTTTCGCAGTAGTTCGTCTTTA
	Neospora caninum	Conventional PCR	NS2	CATGTGGATATTTTGCA	146	[42]
	Neospora caninum	Conventional PCR	NR1	AAACTCCTGGAAGTTAAAG	146	[42]

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