بررسي فنوتايپي و ژنوتايپي تشكيل بيوفيلم در ميان سويه هاي استافيلوكوكوس اورئوس جدا شده از بيماران مبتلا به عفونت ادراري در يك بيمارستان در اصفهان در سال 1395

Phenotypic and Genotypic Characterization of Biofilm Formation among Staphylococcus aureus Strains Isolated from Patients with Urinary Tract Infection in a Hospital in Isfahan during 2016

ﺳﺎﺑﻘﻪ و ﻫﺪف: اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس ﺑﺎﻋﺚ اﯾﺠﺎد ﻃﯿﻒ ﮔﺴﺘﺮده اي از ﻋﻔﻮﻧﺘﻬﺎ در اﻧﺴﺎن از ﻗﺒﯿﻞ ﻋﻔﻮﻧﺖ دﺳﺘﮕﺎه ادراري ﻣﯽ ﺷﻮد ﮐﻪ ﻧﺎﺷﯽ از ﺗﻮاﻧﺎﯾﯽ اﯾﻦ ﺑﺎﮐﺘﺮي در ﺗﺸﮑﯿﻞ ﺑﯿﻮﻓﯿﻠﻢ در ﻧﻘﺎط ﻣﺨﺘﻠﻒ دﺳﺘﮕﺎه ادراري اﺳﺖ. اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﺑﺎ ﻫﺪف ﺗﻌﯿﯿﻦ ﻓﻨﻮﺗﺎﯾﭙﯽ و ژﻧﻮﺗﺎﯾﭙﯽ ﺗﺸﮑﯿﻞ ﺑﯿﻮﻓﯿﻠﻢ در ﻣﯿﺎن ﺳﻮﯾﻪ ﻫﺎي اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس ﺟﺪا ﺷﺪه از ﺑﯿﻤﺎران ﻣﺒﺘﻼ ﺑﻪ ﻋﻔﻮﻧﺖ ادراري در اﺻﻔﻬﺎن اﻧﺠﺎم ﮔﺮﻓﺘﻪ اﺳﺖ. روش ﮐﺎر: در ﻃﯽ ﺳﺎل 1395، در ﻣﺠﻤﻮع 96 ﺟﺪاﯾﻪ اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس از ﯾﮏ ﺑﯿﻤﺎرﺳﺘﺎن ﻣﺮﺟﻊ در اﺻﻔﻬﺎن ﺟﻤﻊ آوري ﺷﺪﻧﺪ و ﺑﺎ اﺳﺘﻔﺎده از ﭘﺮاﯾﻤﺮﻫﺎي اﺧﺘﺼﺎﺻﯽ ﺗﺎ ﺣﺪ ﮔﻮﻧﻪ ﺷﻨﺎﺳﺎﯾﯽ ﮔﺮدﯾﺪﻧﺪ. ﺑﻪ ﻣﻨﻈﻮر ﺑﺮرﺳﯽ ﺗﻮاﻧﺎﯾﯽ ﺗﺸﮑﯿﻞ ﺑﯿﻮﻓﯿﻠﻢ در ﻣﯿﺎن ﺳﻮﯾﻪ ﻫﺎ از روﺷﻬﺎي ﮐﯿﻔﯽ و ﮐﻤﯽ ﻓﻨﻮﺗﺎﯾﭙﯽ ژﻟﻮز ﻗﺮﻣﺰ ﮐﻨﮕﻮ و ﻣﯿﮑﺮوﺗﯿﺘﺮ ﭘﻠﯿﺖ اﺳﺘﻔﺎده ﮔﺮدﯾﺪ و ﺣﻀﻮر ژﻧﻬﺎي ﻣﺨﺘﻠﻒ ﻣﺆﺛﺮ ﺑﺮ ﺗﺸﮑﯿﻞ ﺑﯿﻮﻓﯿﻠﻢ در ﻣﯿﺎن ﺳﻮﯾﻪ ﻫﺎ ﺑﺎ اﺳﺘﻔﺎده از ﭘﺮاﯾﻤﺮﻫﺎي اﺧﺘﺼﺎﺻﯽ ﺗﻌﯿﯿﻦ ﮔﺮدﯾﺪ. ﯾﺎﻓﺘﻪ ﻫﺎ: ﺑﺎ اﺳﺘﻔﺎده از آزﻣﻮن PCR ﺗﻤﺎﻣﯽ ﺟﺪاﯾﻪ ﻫﺎ ﺑﻪ ﻋﻨﻮان ﺳﻮﯾﻪ ﻫﺎي اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس ﻣﻮرد ﺗﺄﯾﯿﺪ ﻗﺮار ﮔﺮﻓﺘﻨﺪ. در ﻣﺠﻤﻮع 5 درﺻﺪ و 72 درﺻﺪ ﺳﻮﯾﻪ ﻫﺎ ﺑﻪ ﺗﺮﺗﯿﺐ واﺟﺪ ﮐﻠﻨﯿﻬﺎي ﺳﯿﺎه رﻧﮓ و ﻗﺮﻣﺰ ﺗﯿﺮه ﺑﻮدﻧﺪ. ﻧﯿﺰ 72 درﺻﺪ ﺳﻮﯾﻪ ﻫﺎ ﺑﻪ ﻋﻨﻮان ﺑﯿﻮﻓﯿﻠﻢ ﻣﺜﺒﺖ ﺷﻨﺎﺳﺎﯾﯽ ﺷﺪﻧﺪ. ﻓﺮاواﻧﯽ ژﻧﻬﺎي fnbA ، clfA ، icaD ، icaA و cna ﻧﯿﺰ در ﻣﯿﺎن ﺳﻮﯾﻪ ﻫﺎي اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس ﻣﻮﻟﺪ ﺑﯿﻮﻓﯿﻠﻢ ﺑﻪ ﺗﺮﺗﯿﺐ 61 ،75 ،86 ،84 و 43 درﺻﺪ ﺑﻮد. ﻧﺘﯿﺠﻪ ﮔﯿﺮي: ﻧﺘﺎﯾﺞ اﯾﻦ ﻣﻄﺎﻟﻌﻪ ﻧﺸﺎن دﻫﻨﺪه ﺷﯿﻮع ﺑﺎﻻي ﺳﻮﯾﻪ ﻫﺎي اﺳﺘﺎﻓﯿﻠﻮﮐﻮﮐﻮس اورﺋﻮس ﻣﻮﻟﺪ ﺑﯿﻮﻓﯿﻠﻢ در ﻣﯿﺎن ﺑﯿﻤﺎران ﻣﺒﺘﻼ ﺑﻪ ﻋﻔﻮﻧﺖ ادراري در ﺑﯿﻤﺎرﺳﺘﺎن ﻣﻮرد ﻧﻈﺮ اﺳﺖ. ﻫﻤﭽﻨﯿﻦ، اﯾﻦ ﯾﺎﻓﺘﻪ ﻫﺎ ﻧﺸﺎن دﻫﻨﺪه اﻫﻤﯿﺖ روش ژﻧﻮﺗﺎﯾﭙﯽ در ﻣﻘﺎﯾﺴﻪ ﺑﺎ روَﻫﺎي ﻓﻨﻮﺗﺎﯾﭙﯽ ﺟﻬﺖ ﺷﻨﺎﺳﺎﯾﯽ ﺳﻮﯾﻪ ﻫﺎي ﻣﻮﻟﺪ ﺑﯿﻮﻓﯿﻠﻢ اﺳﺖ.

Background and objective Staphylococcus aureus is one of the most common causes of a variety of infections in humans, such as urinary tract infection; which is due to its ability to form biofilm in different parts of the urinary tract. In this study we characterized the phenotypic and genotypic analysis of biofilm formation among S. aureus strains isolated from urinary tract infections in Isfahan. Materials and methods During 2016, a total of 96 S. aureus strains were collected from a referral hospital in Isfahan and were identified at the species level using specific primers. The ability of strains to form biofilm was measured using phenotypic qualitative Congo-red agar and quantitative microtiter plate assays. The different genes involved in biofilm formation were detected by the specific primers. Results All isolates were confirmed as S. aureus using the PCR test. The results of Congo-red agar test revealed that 5% and 72% of the strains were able to produce black and dark red colonies, respectively. Moreover, in microtiter plate assay, 72% of the strains were biofilm positive. The frequency of icaA, icaD, clfA, fnbA and cna genes among biofilm producing strains were 84, 86, 75, 61 and 43%, respectively. Conclusion The results of this study indicating the high prevalence of biofilms producing S. aureus strains in the desired hospital. Moreover, our findings revealed the importance of genotypic method for identification of biofilm positive strains compared to phenotypic methods.