Genetic analysis challenges the presence of Ixodes inopinatus in Central Europe: development of a multiplex PCR to distinguish I. inopinatus from I. ricinus

Table 1 Primers used in this study

Gene	Primer name	Sequence (5′—> 3′)	Product size	T_a	References
TROSPA—multiplex	Trospa Iric_F1	GTAAACATCGGCCTAATGG	362 bp	52 °C	This study
	Trospa Iric_R2	GGAAAAAATAATGTTAAAACACC	362 bp
	Trospa Iino_F2	GTTGTTCACAGCGAATACT	233 bp
	Trospa Iino_R1	GAAAAAAATATTAGAACATTAACACTC	233 bp
TROSPA	Trospa-F2	TATGGACACGGCGTCGCTGTC	824 bp/670 bp^b	65 °C	Noureddine et al. [5]
TROSPA	Trospa-R2	GCCCAAGCGCATAAATAAGAAGCGG	824 bp/670 bp^b	65 °C	Noureddine et al. [5]
ITS2	58SRSF3	CTCTTTGAACGCACATTGCGGCCT	831 bp	62 °C	Fukunaga et al. [32]
ITS2	28SRLR2^a	TCTCGCCTGATGTGAGGTCGA	831 bp	62 °C	Fukunaga et al. [32]
Calreticulin	Cal_Fw1	CCCAAGGTGTACCTCAAGG	1238 bp	61.5 °C	This study
Calreticulin	Cal_Rev1	TCCTCTTTATCCTTCTTCTCCG	1238 bp	61.5 °C	This study
16S rRNA	16S-F	TTAAATTGCTGTRGTATT	455 bp	52 °C	Lv et al., [55]
16S rRNA	16S-R1	CCGGTCTGAACTCASAWC	455 bp	52 °C	Lv et al., [55]
COI	COI-F	GAATTAGGACAACCAGGA	1397 bp	55 °C	Noureddine et al. [5]
COI	COI-R	AAAGTATGCTCAGAAGGG	1397 bp	55 °C	Noureddine et al. [5]

ISSN: 1756-3305