Molecular characterization and risk analysis of Giardia duodenalis assemblages in corticosteroid-treated and non-treated patients in Ismailia, Arab Republic of Egypt

Table 1 PCR analysis for the amplification of tpi, bg, and gdh genes of Giardia duodenalis

Gene/Locus	PCR type	Primers code	Primers structure	PCR system*		Final PCR Product	Ref.
Tpi	1^ry	AL3543	5'-AAATIATGCCTGCTCGTCG-3'	Initial denaturation	95 °C / 5 min	530 bp	[34]
	1^ry	AL3544	5'-CAAACCTTITCCGCAAACC-3'	40 cycles	95 °C / 45 s
	2^ry	AL3544	5'-CCCTTCATCGGIGGTAACTT-3'		50 °C / 1 min
		AL3545	5'-GTGGCCACCACICCCGTGCC-3'		72 °C / 1 min
		AL3545	5'-GTGGCCACCACICCCGTGCC-3'	Final extension	72 °C / 7 min
Bg	1^ry	G7F	5’-AAGCCCGACGACCTCACCCGCAGTGC-3’	Initial denaturation	95 °C / 5 min	511 bp	[35, 36]
		G759R	5’-GAGGCCGCCCTGGATCTTCGAGACGAC-3’	40 cycles	95 °C / 30 s
					63 °C / 1 min
					72 °C / 1 min
				Final extension	72 °C / 7 min
	2^ry	GBF	5’-GAACGAACGAGATCGAGGTCCG-3’	Initial denaturation	95 °C / 15 min
		GBR	5’-CTCGACGAGCTTCGTGTT-3’	40 cycles	95 °C / 30 s
					55 °C / 1 min
					72 °C / 1 min
				Final extension	72 °C / 7 min
Gdh	1^ry	GDHeF	5’-TCAACGTYAAYCGYGGYTTCCGT-3’	Initial denaturation	95 °C / 5 min	432 bp	[37, 38]
	1^ry	GDHiR	5’-GTTRTCCTTGCACATCTCC-3’	40 cycles	95 °C / 30 s
	Semi-nested	GDHiF	5’-CAGTACAACTCYGCTCTCGG-3’		56 °C / 1 min
		GDHiR	5’-GTTRTCCTTGCACATCTCC-3’		72 °C / 1 min
		GDHiR	5’-GTTRTCCTTGCACATCTCC-3’	Final extension	72 °C / 7 min

^*The PCR systems have been adjusted to boost the PCR yield of some weak bands. Tpi: Triosephosphate isomerase; bg: β-giardin; gdh: Glutamate dehydrogenase; 1^ry: Primary; 2^ry: Secondary; Ref.: Reference; min: Minutes; s: Seconds; bp: Base pairs

ISSN: 1757-4749