Phenotypic, molecular detection, and Antibiotic Resistance Profile (MDR and XDR) of Aeromonas hydrophila isolated from Farmed Tilapia zillii and Mugil cephalus

Ayoub, Hala F.; khafagy, Ahmed R.; Esawy, Aboelkair M.; El-moaty, Noura Abo; Alwutayd, Khairiah Mubarak; Mansour, Abdallah Tageldein; Ibrahim, Reham A.; Abdel-moneam, Dalia A.; El-Tarabili, Reham M.

doi:10.1186/s12917-024-03942-y

Table 1 List of used A.hydrophila oligonucleotide primers

From: Phenotypic, molecular detection, and Antibiotic Resistance Profile (MDR and XDR) of Aeromonas hydrophila isolated from Farmed Tilapia zillii and Mugil cephalus

Target Gene		Primer sequence (5’-3’)	Amplicon size (bp)	Cycling conditions (35 cycles)			References
Target Gene		Primer sequence (5’-3’)	Amplicon size (bp)	Denaturation	Annealing	extension	References
Virulent genes
Confirmatory gene	16SrRNA	F: AGAGTT TGATCCTGGCTCAG R: GGTTACCTTGTTACGACTT	1200	94 ℃ 1 min	55 ℃ 1 min	72 ℃ 2 min	[63]
Aerolysin	aerA	F: CACAGCCAATATGTCGGTGAG R: GTCACCTTCTCGCTCAGGC	326	94 ℃ 30 s	52 ℃ 30 s	72 ℃ 30 s	[64]
Serine protease	ser	F:ACGGATGCGTTCTTTACTCCA R:CCGTTCATCACACCGTTGTAGTCG	211	94 °C for 1 min	64 °C for 30 s	72 °C for 45 s	[65]
Heat-labile Cytotonic enterotoxin	alt	F: TGACCCAGTCCTGGCACGGC R: GGTGATCGATCACCACCAGC	442	94 ℃ 30 s	55 ℃ 40 s	72 ℃ 45 s	[31]
Heat-stable Cytotonic enterotoxin	ast	F: TCTCCATGCTTCCCTTCCACT R:GTGTAGGGATTGAAGAGCCG	331
Cytotoxic enterotoxin	act	F:AGAAGGTGACCACCACCAAGAACA R:AACTGACATCGGCCTGAACTC	232
Haemolysin	hlyA	F: GGCCGGTGGCCCGAAGATACGGG R:GGCGGCGCCGGACGAGACGGGG	592	95 °C for 2 min	55 °C for 1 min	72 °C for 1 min	[66]
Nuclease	nuc	F:CAGGATCTGAACCGCCTCTATCAGG R:GTCCCAAGCTTCGAACAGTTTACGC	504	94 ºC for 1 min	64 ºC for 30 s	72 ºC for 45 s	[65]
Adhesion	aha	F:GGTATTGTATCCCGGCTCTGTT R:CGGTCCATCGTCGTCCATCTTG	1082	94 ºC for 30 s	60.4 ℃ for 30 s	72 ℃ for 45 s	[67]
Antimicrobial Resistance genes
β-lactamase	bla_pse1	F: ACC GTATTG AGC CTG ATT R: ATTGAA GCC TGT GTT TGA GCTA	321	96 ℃ 30 s	60 ℃ 30 s	72 ℃ 30 s	[68]
β-lactamase	bla_SHV	F: AGGATTGACTGCCTTTTTG R: ATTTGCTGATTTCGCTCG	392	94 ℃ 30 s	54 ℃ 40 s	72 ℃ 40 s	[69]
Sulfonamide	sul1	F: CGCACCGGAAACATCGCTGCAC R:TGAAGTTCCGCCCAAGGCTCG	163	95 ℃ 15 s	65 ℃ 30 s	72 ℃ 30 s	[70]
Tetracycline	tetA	F: GCTACATCCTGCTTGCCTT R: CATAGATCGCCGTGAAGAGG	210	95 ℃ 15 s	60 ℃ 30 s	72 ℃ 30 s	[71]

*Initial denaturation of 5 min at 94 ℃. * Final extension at 72 ℃ extended by 10 min

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ISSN: 1746-6148

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