High Level Aminoglycoside Resistance And Distribution Of The Resistance Genes In Enterococcus faecalis And Enterococcus faecium From Teaching Hospital In Malaysia

Figures & data

Table 1 Primers Used In PCR Assay For 16s rRNA And Detection Of Aminoglycosides Resistance Genes^9,12

Genes	Primer Sequences (5’ – 3’)	Product Size (bp)
16s rRNA	F: GTGCTGCAGAGAGTTGATCCTGGCTCAG R: CACGATCCTACGGGTACCTTGTTACGACTT	1465
aac(6’)-Ie-aph(2”)-Ia	F: CAGGAATTTATCGAAAATGGTAGAAAAG R: CACAATCGACTAAAGAGTACCAATC	369
aph(2ꞌꞌ)-Ib	F: CTTGGACGCTGAGATATATGAGCAC R: GTTTGTAGCAATTCAGAAACACCCTT	867
aph(2ꞌꞌ)-Ic	F: CCACAATGATAATGACTCAGTTCCC R: CCACAGCTTCCGATAGCAAGAG	444
aph(2ꞌꞌ)-Id	F: GTGGTTTTTACAGGAATGCCATC R: CCCTCTTCATACCAATCCATATAACC	641
aph(3ꞌ)-IIIa	F: GGCTAAAATGAGAATATCACCGG R: CTTTAAAAAATCATACACGCTCGCG	523

Table 2 Distribution Of Enterococcus Species

Specimen	E. faecalis (n=50), n (%)	E. faecium (n=25), n (%)	Total Isolates (n=75), n (%)
Pus	29 (38.7)	9 (12)	38 (50)
Blood	14 (18.6)	10 (13.3)	24 (32)
Urine	2 (2.7)	6 (8)	8 (11)
Others	5 (6.7)	0 (0)	5 (7)
Total	50 (66.7)	25 (33.3)	75 (100)

Abbreviation: n, number of occurrence.

Figure 1 Antibiotic resistance profile of E. faecalis and E. faecium.

Abbreviations: CN, gentamicin; S, streptomycin; AMP, ampicillin; VA, vancomycin; E, erythromycin; C, chloramphenicol; TE, tetracycline; LZD, linezolid.

Table 3 Antimicrobial Patterns Of E. faecalis Isolates

Antibiotic Class	Antibiotic Agents	Susceptible, n (%)	Resistance, n (%)
Aminoglycosides	Gentamicin 120 µg	26 (52)	24 (48)
	Streptomycin 300 µg	27 (54)	23 (46)
Beta-lactams	Ampicillin 30 µg	32 (76)	12 (24)
Glycopeptides	Vancomycin 10 µg	47 (94)	3 (6)
Macrolides	Erythromycin 15 µg	2 (4)	48 (96)
Chloramphenicol	Chloramphenicol 30 µg	27 (54)	23 (46)
Tetracyclines	Tetracycline 30 µg	1 (2)	49 (98)
Oxazollidinones	Linezolid 30 µg	48 (96)	2 (4)

Note: Antimicrobial breakpoints were interpreted according to CLSI (2016) guidelines.

Table 4 Antimicrobial Patterns Of E. faecium Isolates

Antibiotic Class	Antibiotic Agents	Susceptible, n (%)	Resistance, n (%)
Aminoglycosides	Gentamicin 120 µg	4 (16)	21 (84)
	Streptomycin 300 µg	8 (32)	17 (68)
Beta-lactams	Ampicillin 30 µg	4 (16)	21 (84)
Glycopeptides	Vancomycin 10 µg	25 (100)	0 (0)
Macrolides	Erythromycin 15 µg	0 (0)	25 (100)
Chloramphenicol	Chloramphenicol 30 µg	17 (68)	8 (32)
Tetracyclines	Tetracycline 30 µg	0 (0)	25 (100)
Oxazollidinones	Linezolid 30 µg	25 (100)	0 (0)

Note: Antimicrobial breakpoints were interpreted according to CLSI (2016) guidelines.

Figure 2 (A) Representative image of the amplified PCR products of HLGR gene, aac(6’)-Ie aph(2”)-Ia of Enterococcus isolates (expected size: 369 bp). (B) Representative image of the amplified PCR products of HLSR gene, aph(3’)-IIIa of Enterococcus isolates (expected size: 523 bp).

Abbreviations: Lane M, 100 bp ladder; NC, negative control (DNA-free template); PC, positive control (ATCC strain 51299); Lanes 1–21, Enterococcus isolates.

Shente V, Grover N, Kumar M. Analysis of aminoglycoside modifying enzyme genes responsible for high-level aminoglycoside resistance among enterococcal isolates. J Pathog. 2017;2017:325695229435370

 PubMedGoogle Scholar

Vakulenko SB, Donabedian SM, Voskresenskiy AM, et al. Multiplex PCR for detection of aminoglycoside resistance genes in enterococci. Antimicrob Agents Chemother. 2003;47(4):1423–1426. doi:10.1128/aac.47.4.1423-1426.200312654683

 PubMed Web of Science ®Google Scholar