Detection of Antibiotic-Resistant Gene from Isolates of Diabetic Ulcer Patients
DOI:
https://doi.org/10.26418/ijeasr.v1i1.40988Abstract
Antibiotic resistance can be caused by the presence of several genes encoding antibiotic resistance found in certain bacteria. S. aureus bacteria contained blaZ (97.4%), mecA (42.3%) and tetM genes (20.2%). Bacillus sp. contained blm (87.2%), tetL (44%) and tetB genes (2.9%). E. coli bacteria contained tem (80.9%)dan shv genes (14.28%). Clostridium spp. Bacteria contained cat gene (36%), as well as S. pyogens contained ermT (36.5%). Specific primers made in silico based on bioinformatics were then analyzed by PCR (Polymerase Chain Reaction) and electrophoresis. The purpose of this study was to detect the presence of blaZ, mecA, tetM, blM, tetL, tetB, tem, shv, cat, and ermT genes. The research began by searching for the nucleotide sequences for the ermT, cat, tetB, and blaZ genes through the Primer 3Plus program and analyzed using OligoAnalyzer 3.1. In vitro detection was carried out by extracting DNA and then amplification using PCR, and analyzed by agarose gel electrophoresis. The results of the analysis of the primer criteria for the ermT gene and the blaZ gene at the optimum Tm 55 ° C, cat gene and the tetB gene at the optimum Tm 60 ° C have met all the primer criteria. The results obtained are mecA gene (533 bp), tetM gene (366 bp), blm gene (1107 bp), tetL gene (267 bp), tetB gene (186 bp), tem gene (1073 bp), cat gene (164 bp), the ermT gene (202 bp) has been detected successfully, while blaZ and shv have not been detected successfully.References
Kannan I, Premavathy KR, Sambandam C. Isolation and Antibiotic Susceptibility of Bacteria from Foot Infections. Int J Res Med Sci. 2014: 2(2); 457-461
Chellakkannu M, Revathi V, Panneerselvam T. Bacteriological Study of Diabetic Foot on the Antibiotic and Topikal Ointment Therapy. Journal of Pharmaceutical and Life Sciences. 2016; 2: 259-266.
Jamali H, Paydarb M, Radmehrc B, Salmah I and Dadrasniaa A, 2015. Prevalence and antimicrobial resistance of Staphylococcus aureus isolated from raw milk and dairy products. Food Control, 54: 383388
Tahmasebi H, Talebi R, Zarif BR, Isolated of Bacillus Cereus in Chicken Meat and Investigation β-Lactamase Antibiotic-Resistant in Bacillus Cereus from Chicken Meat. Ad. In Life. Sci. 2014: 4(4); 200-206.
Saeed BMS, Abbas BA, Al-Jaadan SAN. Molecular Detection Of Tetracycline Resistance Genes In Bacillus cereus Isolated From Food Sources. Basrah Journal of Veterinary Research. 2018 ; 17(3) : 1-8.
Jiang X, Shi L. Distribution Of Tetracycline and Trimethoprim/SUlfamethoxazole Resistance Genes in Aerobic Bacteria Isolated From Cooked Meat Products in Guangzhou,China. Journal Food Control. 2013; 30(1): 30-34
Michael NS, Saadi AT. Detection of bla CTX-M, bla TEM-01 and bla SHV Genes in Multidrug Resistant Uropathogenic E. coli Isolated from Patients wits Recurrent Urinary Tract Infections. International Journal of Medical Research & Health Sciences. 2018: 7(9); 81-89.
Nurmala, Virgiandhy IGN, Andriani, Liana DF. Resistensi dan sensitivitas bakteri terhadap antibiotik di RSU dr. Soedarso Pontianak tahun 2011-2013. Fakultas Kedokteran Universitas Tanjungpura. 2015;3(1) : 21-28
Huang et al.Type B Chloramphenicol Acetyltransferases Are Responsible for Chloramphenicol Resistance in Riemerella anatipestifer, China. Journal Frontiers in Microbiology. 2017 ; 8 : 297
Woo YCP, Amanda TCP, Lau KPS, Fung YMA, Yuen K. Phenotypic and Molecular Characterization of Erythromycin Resistance in Four Isolates of Streptococcus-Like Gram-Positive Cocci Causing Bacteremia. Journal Clinical Microbiology. 2004 ; 42(7) :3303-3305
Borah P. Primer Designing for PCR. Sci Vis. 2011;11(3):134–6.
Muladno. Teknologi Rekayasa Genetika. 2nd ed. Bogor: IPB Press; 2010.
Jusuf M. Genetika I: Struktur dan Ekspresi gen. 1st ed. Jakarta: Sagung Seto; 2001.
Yusuf Z. Polymerase Chain Reaction (PCR). Saintek. 2010;5(6):1–6.
Maksum IP, Amalia R, Kamara DS, Diana S, Rachman RW. PCR Multipleks untuk Identifikasi Mycobacterium tuberculosis Resisten terhadap Isoniazid dan Rifampisin pada Galur Lokal Balai Laboratorium Kesehatan Provinsi Jawa Barat Multiplex PCR for Identification the Isoniazid- and Rifampin-resistant Mycobacterium tuberculosis (Local Strain of Balai Laboratorium Kesehatan Provinsi Jawa Barat) 2018;4(Nov):107–18.
Borah P. Primer Designing for PCR. Sci Vis. 2011;11(3):134–6.
Sasmito D, Kurniawan R, Muhimmah I. Karakteristik primer pada polymerase chain raction (PCR) untuk sekuensing DNA: mini review. SNI Med. 2014;5:93–102.
Budiani A, Putranto RA, Santoso D. Kloning gen penyandi β-1 , 6-glukanase kapang secara cepat dengan teknik RT-PCR menggunakan primer spesifik. Menara Perkeb. 2009;77(1):45–5
