A Two-Tube Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Viral and Bacterial Pathogens of Infectious Diarrhea

Table 3

Primer sequences and product sizes used in this study.

Pathogen	Target	Sequence 5′-3′	Size	References
Shigella and EIEC	ipaH	AGGTGACACTATAGAATA ^aACCATGCTCGCAGAGAAACT	213	[10]
		GTACGACTCACTATAGGGA ^bTCAGTACAGCATGCCATGGT
Vibrio parahaemolyticus	tlh	AGGTGACACTATAGAATAACTCAACACAAGAAGAGATCGACAA	246	[30]
		GTACGACTCACTATAGGGAGATGAGCGGTTGATGTCCAA
EHEC and EPEC	eaeA	AGGTGACACTATAGAATAAGGTCGTCGTGTCTGCTA	293	[31]
		GTACGACTCACTATAGGGACCGTGGTTGCTTGCGTTTG
Salmonella	invA	AGGTGACACTATAGAATAGTGAAATTATCGCCACGTTCGGGCAA	323	[32]
		GTACGACTCACTATAGGGATCATCGCACCGTCAAAGGAACC
Yersinia	ail	AGGTGACACTATAGAATATAATGTGTACGCTGCGAG	389	[5]
		GTACGACTCACTATAGGGAGACGTCTTACTTGCACTG
Vibrio cholerae	ctx	AGGTGACACTATAGAATAACAGTAACTTAGATATTGCTCCAG	507	This study
		GTACGACTCACTATAGGGAACCATTCTTAAAAGTAATGATAGCCA
Campylobacter jejuni	mapA	AGGTGACACTATAGAATACTATTTTATTTTTGAGTGCTTGTG	627	[33]
		GTACGACTCACTATAGGGAGCTTTATTTGCCATTTGTTTTATTA
Noroviruses GI	RDRP	AGGTGACACTATAGAATACGCTGGATGCGCTTCCATGA	162	[21]
		GTACGACTCACTATAGGGAGCAAGAGGGTCAGAAGCATT
Rotavirus	VP6	AGGTGACACTATAGAATAAAGTCTTCCACATGGAGGT	210	Modified [21]
		GTACGACTCACTATAGGGAARRTTICCAATTCCTCCAGT
Norovirus GII	RDRP	AGGTGACACTATAGAATACAGACAAGAGCCAATGTTCA	279	Modified [21]
		GTACGACTCACTATAGGGATTTCTAATCCAGGGGTCAATT
Human astrovirus	ORF1a	AGGTGACACTATAGAATACGTCATTATTTGTTGTCATA	326	[21]
		GTACGACTCACTATAGGGACATGTGCTGCTGTTACTATG
Enteric adenovirus	Hexon	AGGTGACACTATAGAATATGTACAAGCCAGNTGTAGCTC	388	This study
		GTACGACTCACTATAGGGAAAGCAGTAATTTGGCANTTCGT
Human bocavirus 1^c	VP1	AGGTGACACTATAGAATAAAACCCATCACTCTCAATGCTT	412	This study
		GTACGACTCACTATAGGGACAGTATGTCTTCTTTCTGGACG
Human bocavirus 2	VP1	AGGTGACACTATAGAATAAAATCCACCACTATCCATGCTC	412	This study
		GTACGACTCACTATAGGGACGGTGTGTCTTCTTTCTGGTCT

aUniversal primers-F: AGGTGACACTATAGAATA; ^buniversal primers-R: GTACGACTCACTATAGGGA; ^cthe primers for HBoV1 and HBoV2 are equally mixed, the amplicon sizes of both PCR products are exactly the same, and the primers are located at the same position in the corresponding viral genome; this method is able to identify the presence of HBoV1 and HBoV2 but cannot classify the subtypes.