Table 1 The primers and reactions condition applied to detect enteric pathogens in this study

EPEC

eae

CCACGGTTTATCAAACTGATAACG

105

Each stool specimen was inoculated to MAC media to culture DEC at 37 °C for 18 h, And then ten putative DEC colonies were selected to mix with 150 μL water to extract DNA at 100 °C for 10 min, and then the 20 μL volume of qPCR system is composed of 10 μL master mix (Takara Bio Inc, Shiga, Japan), 1 μL forward primer (10 μmol), 1 μL reverse primer (10 μmol), 1 μL DNA template and 7 μL H₂O. The cycling conditions for each subtype DEC was 95 °C for 5 min, 40 cycles of 95 °C for 5 s, 60 °C for 30 s. The fluorescence recorded was at the annealing stage

[20, 21]

EHEC

stx1

ACTTCTCGACTGCAAAGACGTATG

132

ACAAATTATCCCCTGAGCCACTATC

stx2

CCACATCGGTGTCTGTTATTAACC

93

GGTCAAAACGCGCCTGATAG

ETEC

elt

TTCCCACCGGATCACCAA

62

CAACCTTGTGGTGCATGATGA

estA

CCTTTCGCTCAGGATGCTAAAC

128

CAGTAATTGCTACTATTCATGCTTTCAG

estB

CTTTCCCCTCTTTTAGTCAGTCAACT

137

GCAGTAAAATGTGTTGTTCATATTTTCTG

EAEC

aggR

CAGCGATACATTAAGACGCCTAAAG

116

CGTCAGCATCAGCTACAATTATTCC

EIEC

ipaH

ACCATGCTCGCAGAGAAACT

175

TCAGTACAGCATGCCATGGT

RVA

VP6

GACGGVGCRACTACATGGT

382

RVA, NoV GI, NoV GII, SaV and As were RNA viruses, complementary DNA (cDNA) was synthesized using a random primer (Takara Bio Inc, Shiga, Japan) at 55 °C for 1.5 h, followed by 100 °C for 10 min, and holding at 4 °C. The reaction condition of RVA was 94 °C for 5 min, followed by 40 cycles at 94 °C for 1 min, 42 °C for 1 min, 72 °C for 1 min, and with final extension at 72 °C for 10 min. Multiplex RT-PCR was used to detect the presence of NoV GI, NoV GII, and SaV, the thermal profile consisted of 94 °C for 5 min, 40 cycles of 94 °C for 70 s, 49 °C for 70 s, and 72 °C for 1 min, followed by 72 °C for 10 min. The thermal profile of As was 94 °C for 5 min, 40 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min, followed by 72 °C for 10 min

[22]

GTCCAATTCATNCCTGGTGG

NoV GI

NoV GII

SaV

Polymerase

TGACGATTTCATCATCACCATA

331/319

[23]

TGACGATTTCATCATCCCCGTA

GATTACTCCAGGTGGGACTCCAC

GATTACTCCAGGTGGGACTCAAC

GATTACTCCAGGTGGGATTCAAC

GATTACTCCAGGTGGGATTCCAC

As

Capsid

CAACTCAGGAAACAGGGTGT

449

[24]

TCAGATGCATTGTCATTGGT

Ad

Hexon

TTCCCCATGGCICAYAACAC

482

The thermal profile was 94 °C for 5 min, 40 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min, followed by 72 °C for 10 min

[25]

CCCTGGTAKCCRATRTTGTA

Blastocistis hominis

SSU-rRNA

CGAATGGCTCATTATATCAGTT

260

The thermal profile was 94 °C for 5 min, 40 cycles of 94 °C for 30 s, 53 °C for 30 s, and 72 °C for 1 min, followed by 72 °C for 10 min

[26]

TCTTCGTTACCCGTTACTGC

Cryptosporidium spp.

18S-rRNA

TTCTAGAGCTAATACATGCG

The primary cycle consisted of 94 °C for 5 min, 35 cycles of 94 °C for 50 s, 55 °C for 1 min and 72 °C for 90 s, followed by 72 °C for 10 min, the annealing step for a second reaction was 58 °C

[27]

CCCATTTCCTTCGAAACAGGA

GGAAGGGTTGTATTTATTAGATAAAG

840

CTCATAAGGTGCTGAAGGAGTA

Giardia lamblia

Tim

AAATIATGCCTGCTCGTCG

The thermal profile of first round was 94 °C for 1 min, 53 °C for 1 min, and 72 °C for 1 min, followed by 72 °C for 10 min. A second reaction was carried out similarly

[28]

CAAACCTTITCCGCAAACC

CCCTTCATCGGIGGTAACTT

530

GTGGCCACCACICCCGTGCC