Table 1. Established combination of primers and probes using the multiplex real-time PCR assay for the major parasites causing gastroenteritis.

Target organism	Forward primer sequence (5'-3')	Reverse primer sequence (5'-3')	Optimized probe sequence (5'-3')	Target	Accession number of target sequence
Gymnophalloides seoi	AGTGTTGTTTGGGCTCATCA	AACCTTAATCGGTGGGAACA	FAM-TGTTTATGGTTGGTTGATGTTAAGACTTCGGT-black hole quencher 1	CO1	AF096234.3
Blastocystis hominis	GGAGAGGGAGCCTGAGAGAT	AACATTGTTCCGCATTGTGA	VIC-ATCCTGACACAGGGAGGTAGTG-non fluorescent quencher MGB	18s rRNA	EU482085.1
Cryptosporidium parvum	TGTGTTCAATATCTCCCTGCAAA	GCATGTCGATTCAATTTGTCA	Quasar670-CCTCCTGGATTCAATTTGTCA-black hole quencher plus	Cowp 1	AB089292.1
Giardia lamblia	GAGGTCAAGAAGTCCGCCG	CAAGGGACTTGCGGAAGTTT	FAM-ACGATCAAGGAGGAGATCGA-black hole quencher plus	beta-giardin	XM_001705373.1
Entamoeba histolytica	GCGGACGGCTCATTATAACA	TGTCGTGGCATCCTAACTCA	VIC-AAATGGCCAATTCATTCAATG-non fluorescent quencher MGB	18s rRNA	X65163.1
Metagonimus yokogawai	CTATGGCGGTTTAGTGTTGGC	ACTGCCGTCTTCAAATCCAG	Quasar670-TGGGCGCATCACATGTTTATGGTG-black hole quencher plus	CO1	AB470519.1
Clonorchis sinensis	GGTGGTTTGAGCTCATCATATGT	CGAGTTCCAGCAAGCATATATAATC	FAM-CGGTTACTATGATTATAGGTGTGCC-black hole quencher 1	CO1	FJ381664.2
Dientamoeba fragilis	TTAGAACCTTAGACAACGGATGTCTTG	TGTGCATTCAAAGATCGAACTTATC	VIC-TGTGATAAGCGGCTAGAATTGC-non fluorescent quencher MGB	18s rRNA	JQ677163.1

CO1 = cytochrome c oxidase subunit 1; Cowp = Cryptosporidium oocyst wall protein