TY  - JOUR
T1  - AGAR WELL DİFÜZYON YÖNTEMİNDE STANDARDİZASYON ÇALIŞMASI
TT  - STANDARDIZATION STUDY FOR AGAR WELL DIFFUSION METHOD
AU  - Aytar, Mehmet
AU  - Oryaşın, Erman
AU  - Başbülbül, Gamze
AU  - Bozdoğan, Bülent
PY  - 2019
DA  - December
JF  - Bartın University International Journal of Natural and Applied Sciences
JO  - JONAS
PB  - Bartın Üniversitesi
WT  - DergiPark
SN  - 2667-5048
SP  - 138
EP  - 145
VL  - 2
IS  - 2
LA  - tr
AB  - Antibiyotikduyarlılık testleri hastalık etkeni mikroorganizmalara karşı kullanılabilecekantibiyotiklerin belirlenmesinde önemlidir. Antibiyogram testlerinde farklıkuruluşlar tarafından standardizasyon çalışması yapılmasına karşın agar well testleri için standardizasyonçalışmasının yapılmadığı görülmüş ve bu çalışmada disk difüzyon testiylekarşılaştırmalı standardizasyon çalışması yapılması amaçlanmıştır. Testler 0,5 McFarlandbulanıklıkta bakteri süspansiyonu ile 3 farklı kalınlıkta (4, 6, 8 mm) Mueller Hintonagar besiyeri ve 3 farklı çapta kuyucuk (4, 6, 8 mm) kullanılarak üç tekrarlıolacak şekilde yürütülmüştür. S. aureus ATCC25923 ve E. coli DH10B bakterileriiçin sırasıyla eritromisin (15µg) ve kloramfenikol (10µg) antibiyotiklerikullanılmıştır. Tüm petriler 16-18 saat, 37°C’de inkübe edilmiştir. İnkübasyonsonunda disklerin ve kuyucukların etraflarında oluşan inhibisyon zonlarıölçülmüştür. Antibiyotik duyarlılık testinde kullanılan agar well yöntemi için yaptığımız standardizasyon çalışmasında enuygun Mueller Hinton Agar besiyeri kalınlığı ve kuyucuk çapı saptanmıştır. Agarkalınlığı ne olursa olsun, disk difüzyon zon çapıyla agar well kuyucuk zon çapının 8 mm kuyucukla eşleştiği görülmüştür.Agar kalınlığı arttıkça inhibisyon zonu artmaktadır. Disk difüzyon testindestandart 4 mm kalınlığındaki besiyeri olduğu için bu kalınlıktaki besiyerindedisk difüzyon sonucuna uyan inhibisyon zonu oluşturan kuyucuk çapı 8 mm olaraksaptanmıştır. Standart ölçüm için 4 mm agar kalınlığı ve 8 mm kuyucuk çapıolmalıdır.Çalışmamızagar well difüzyon konusundaki ilkstandardizasyon çalışmasıdır. Diğer laboratuvarların benzer standardizasyonçalışmaları yapmaları halinde agar welldifüzyon yöntemini tüm dünyada geçerli olabilecek bir standardakavuşturabiliriz.
KW  - Agarwell
KW  - Standardizasyon
KW  - Besiyeri kalınlığı
KW  - Kuyucuk çapı
N2  - Antibiotic sensitivity tests are important todetermine antibiotics that can be used against disease-causing microorganisms.Although different standardization studies for disk diffusion and MIC tests wereestablished by several organizations, no standardization study was done for agar well diffusion method.Standardization for disk diffusion method, antibiogram, was done first by WorldHealth Organization (WHO). Today many organizations including, The EuropeanCommittee on Antimicrobial Susceptibility Testing (EUCAST) in Europe, Clinical and Laboratory Standards Institute(CLSI) in USA, and Standardization of Antibiotic Sensitivity Tests (ADTS) groupof Turkish Microbiology Society (TMC) in Turkey developed their ownstandardization methods. TMC advises the use of EUCAST protocols for studies inTurkey.The aim of the present study was standardization of agar well diffusion method. Diskdiffusion method from EUCAST was used as a reference method to compare resultsof agar well diffusion test. For bothdisk diffusion method as well as agarwell diffusion test 0.5 McFarland bacterial suspension and Mueller HintonAgar (MHA) media were used. For agar welldiffusion test 3 different thicknesses, 4 mm, 6 mm and 8 mm, of MH agar wasused. Also three different well diameter, 4 mm, 6 mm and 8 mm were tested. Alltests were performed as three repetitions. Two antibiotics were used for diskdiffusion tests and agar welldiffusion tests. The concentration of erythromycin and chloramphenicol were 15 µgand 10 µg, respectively. A Gram positive and a Gram negative strain were usedfor susceptibility tests, S. aureus ATCC25923 and E. coli DH10B strains,respectively. All of plates were incubated at 37°C for 16-18 hours. At the endof incubation, the inhibition zones around wells were measured. The inhibition zones of S.aureus ATCC 25923with erythromycin disk were 29, 30 and 31 mm on 4, 6 and 8 mm agar,respectively. Agar well diffusionwith 4, 6 and 8 mm MHA media, erythromycin inhibition zones were 21, 25 and 29mm, 22, 26 and 30 mm, and 23, 27 and 31 mm, respectively. The inhibition zones of E. coliDH10B with chloramphenicol disk were 25, 26 and 27 mm on 4, 6 and 8 mm agar,respectively. Agar well diffusionwith 4, 6 and 8 mm MHA media, chloramphenicol inhibition zones were 5, 6 and 7mm, 15, 16 and 17 mm, and 25, 26 and 27 mm, respectively. It was observed thatthe disk diffusion inhibition zones were in accordance with the results of 8 mmagar well zone, in every MH agarthicknesses. As the standard for disk diffusion test the MHA thickness is 4 mm,the agar well diameter for thisthickness is 8 mm. So for standardization of agar well diffusion tests, the MHA thickness should be 4 mm and thediameter should be 8 mm. A standardization method for agar well diffusion test is necessary to compare results obtainedelsewhere. Our study is the first study for standardization of agar well diffusion test.Standardization assays should be done by other laboratories to determine aworldwide usable and comparable agar welldiffusion method.
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UR  - https://dergipark.org.tr/tr/pub/jonas/issue//611508
L1  - https://dergipark.org.tr/tr/download/article-file/889117
ER  -