Pola kepekaan isolat Eschericia coli dari penderita diare di Yogyakarta dan analisa faktor resistensinya

HUDA, Misbahul, Dra. Susi Iravati, Apt.,PhD

2004 | Tesis | S2 Ilmu Kedokteran Tropis

Diare merupakan masalah kesehatan masyarakat di Indonesia, karena masih tingginya angka kesakitan dan kematian. Escherichia coli merupakan penyebab 25% kasus diare di negara-negara berkembang. Pemakaian antibiotik yang tidak teratur pada diare akut dapat memberikan dampak meningkatkan resiko efek samping dan resistensi antibiotik pada bakteri. Sebagian Escherichia coli telah resisten terhadap beberapa jenis antibiotik. Resistensi dapat disandi oleh kromosom atau plasmid. Resistensi ekstrakromosomal disandi oleh plasmid. Plasmid merupakan pembawa sifat resistensi yang dapat menyebabkan resistensi, karena dapat ditularkan ke sel lain maupun ke spesies lain. Penelitian ini bertujuan melihat pola kepekaan dan menganalisa faktor resistensi Escherichia coli yang diisolasi dari penderita diare di Yogyakarta. Sampel feces penderita diare dikumpulkan dari 4 rumah sakit di Yogyakarta yaitu RSPKU Muhammadiyah, RSUD Sleman, RSUP DR. Sardjito dan RSUD Bantul. Isolasi dan identifikasi E.coli dilakukan pada media selektif Mac Conkey Agar pada 37oC semalam dan dilanjutkan uji Biokimia pada KIA, SSS, LIA dan MIO Ureum. Selanjutnya dilakukan uji kepekaan terhadap isolat E.coli dengan metode Difusi cara Kirby Bauer, menggunakan 6 jenis antibiotik yaitu Sulfamethoxazole/Trimethoprim, Chloramphenicol, Amoxicilin, Tetracyclin, Ciprofloxacin, Amoxycillin-Clav-Acid. Selanjutnya dilakukan isolasi plasmid dari semua isolat E.coli yang resisten antibiotik tersebut dengan metode boiling menggunakan buffer TELT, kemudian dilakukan elektroforesis pada agarose 1%. Plasmid hasil isolasi ditransformasikan ke dalam E.coli XL-1 Blue. Kemudian dilakukam isolasi plasmid dari transforman yang dipeoleh. Hasil isolasi plasmid dielektroforesis dan dianalisa. Hasil penelitian menunjukkan bahwa dari 60 sampel feces penderita diare didapatkan 51 isolat E.coli. Pola kepekaan isolat E.coli tersebut menunjukkan bahwa 82,3% resisten terhadap Sulfamethoxazole/ Trimethoprim, 64,7% terhadap Amoxicilin, 52,9% terhadap Amoxycillin-Clav-Acid, 47% terhadap Tetracyclin, 33,3% terhadap Chloramphenicol, dan 7,8%terhadap Ciprofloxacin. Isolat yang masih sensitif terhadap keenam antibiotik tersebut hanya 11,76%, sedangkan yang resisten terhadap 1 jenis antibiotik 11,76% dan yang multidrug resistance 76,47%. Plasmid yang ditransformasikan ke E.coli XL-1 Blue tidak berhasil diisolasi kembali.

Diarrhea is one of the community diseases in Indonesia as implied in the high rate of morbidity and mortality. Escherichia coli has accounted for 25% of acute diarrhea cases in developing countries. Irregular use of antibiotics for acute diarrhea has resulted in higher risk of side effect and bacterial resistance to antibiotics. Some Eschericia coli has developed resistance to some types of antibiotics. The resistance can be encoded by chromosome or plasmid. Extra chromosomal resistance is encoded by plasmid. Plasmid has caused resistance and it may affect other cells or other species. This research aims at identifying the sensitivity pattern and analyzing the resistence of Escherichia coli isolated from the diarrheic patients in Yogyakarta. Feces samples taken from diarrheic patients were collected from 4 hospitals in Yogyakarta including RSPKU Muhammadiyah, RSUD Sleman, RSUP Dr. Sardjito, and RSUD Bantul. E. coli was isolated and identified on the selective media of Mac Conkey Agar at 37°C overnight and biochemical testing at KIS, SSS, LIA, and MIO Ureum. Next, a test of sensitivity to E. coli isalate was performed to by Kirky Bauer’s diffusion method using 6 types of antibiotics: Sulfamethoxazole/Trimethoprim, Chloramphenicol, Amoxicilin, Tetracyclin, Ciprofloxacin, Amoxycyllin-Clav-Acid. Plasmid isolation from all E. coli isolates resistant to antibiotics was performed by buffer TELT boiling method. Electrophoresis at 1% agar was sequentially performed. The plasmid of the isolation was transformed into E.coli XL-1 Blue. Plasmid which was resulted from the transformer was isolated. Electrophoresis and analysis were formed for the result of the plasmid. The results of the study suggested that there were 51 E. coli isolates of the 60 feces sample from the diarrheic patients. The sensitivity pattern of E. coli suggested that 82.5% were resistant to Sulfamethoxazole/Trimethoprim, 64.7% to Amoxicillin, 52.9% TO Amoxycillin-Clav-Acid, 47% to Tetracyclin, 33.3% to chloramphenicol, and 7.8 to Ciprofloxacin. There were only 11% of isolates still sensitive to the six antibiotics and 76.47% of isolates developing resistance to multi-drugs. The plasmid that had been transformed into E. coli could not be re-isolated.

Kata Kunci : Kedokteran Tropis, Epidemiologi Resistensi E Coli, resistence, Escherichia coli, plasmid, E.coli XL-1 Blue.