First molecular detection of Francisella tularensis in turtle (Testudo graeca) and ticks (Hyalomma aegyptium) in Northwest of Iran Author Tukmechi, Amir a.tukmachi@urmia.ac.ir Author Ownagh, Abdulghaffar Author Enferadi, Ahmad Author Khademi, Peyman text International Journal for Parasitology: Parasites and Wildlife 2024 2024-04-30 23 100892 100892 http://dx.doi.org/10.1016/j.ijppaw.2023.11.005 journal article 294515 10.1016/j.ijppaw.2023.11.005 5e440607-6c27-489b-a9ff-c9587c75b59e 2213-2244 PMC10772711 38192304 10933773

Table 1 The PCR technique used for identifying the Francisella genus and subspecies includes specific primers, product sizes and a thermal program, which are described as follows.

Protocol	Primer	Sequence 5’——3’	product size (bp)		PCR condition (cycle)
Name
Normal-	16SrRNA-F	TGGTGTAGCGGTGAAATGCGTA	495		95 ◦C for 4 m, 95 ◦C for 55s, Touchdown 66–63 (5) for 55s, 72 ◦C for 55 m, 72 ◦C for
PCR	16SrRNA-R	GCCTTGTCAGCGGCAGTCTTAATA			7 m. (32)
Nested-PCR	16SrRNA-	TGGTAGTCCACGCTGTAAACGATG	306		95 ◦C for 4 m, 95 ◦C for 15s, 66 ◦C for 15s, 72 ◦C for 7 m. (30)
NF
	16SrRNA-	GCGGGACTTAACCCAACATTTCAC
	NR
Normal-	pdpD -F	TGGGTTATTCAATGGCTCAG	F. tularensis	136	95 ◦C for 4 m, 95 ◦C for 15s, 66 ◦C for 15s, 72 ◦C for 7 m. (30)
PCR			F. holarctica	–
	pdpD-R	TCTTGCACAGCTCCAAGAGT	F. mediasiatica	280
			F. novicida
Normal-	RD1-F	TGATTCTGTACGCTGGCGTTGT	F. holarctica	520	95 ◦C for 4 m, 95 ◦C for 15s, 62 ◦C for 15s, 72 ◦C for 7 m. (30)
PCR	RD1-R	AAGCGCACCGTAGCTTTCATCT