بررسي فراواني ژنهاي حدت اگزوتوكسين A، اگزوآنزيم، آلژينات و oprI وoprL در جدايه‌هاي سودوموناس آئروژينوزا دامي و انساني و تعيين الگوي مقاومت آنتي‌بيوتيكي

Frequency of Exotoxin A, Exoenzyme, Alginate and PprI and PprL Virulence Genes in Animal and Human Pseudomonas Aeruginosa Isolates and Determination of Antibiotic Resistance Pattern

زﻣﻴﻨﻪ و اﻫﺪاف: ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﻳﻨﻮزا ﻣﻬﻤﺘﺮﻳﻦ ﻋﺎﻣﻞ ﻋﻔﻮﻧﺖ ﻫﺎي ﻣﺨﺘﻠﻒ ﺑﻴﻤﺎرﺳﺘﺎﻧﻲ و ورم ﭘﺴﺘﺎن در ﮔﺎوﻫﺎي ﺷـﻴﺮي و اﻳﺠﺎد ﻣﻘﺎوﻣﺖ آﻧﺘﻲﺑﻴﻮﺗﻴﻜﻲ ﻣﺤﺴﻮب ﻣﻲﺷﻮد. ﻫﺪف اﻳﻦ ﻣﻄﺎﻟﻌﻪ ﺗﻌﻴﻴﻦ ﻣﻘﺎوﻣﺖ آﻧﺘﻲﺑﻴﻮﺗﻴﻜﻲ ﺑﺎﻛﺘﺮي ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﻳﻨﻮزا و ﺣﻀﻮر ژﻧﻬﺎيﺣﺪت در ﻧﻤﻮﻧﻪﻫﺎي اﻧﺴﺎﻧﻲ و داﻣﻲ ﺑﻮده اﺳﺖ. ﻣﻮاد و روش ﻛﺎر: در اﻳﻦ ﺗﺤﻘﻴﻖ 102 ﺳﻮﻳﻪ اﻧﺴﺎﻧﻲ و داﻣﻲ از ﺳﻮدوﻣﻮﻧﺎس آﺋﺮوژﻳﻨﻮزا ﻣﻮرد ﺑﺮرﺳﻲ ﻗﺮار ﮔﺮﻓﺖ. در ﺑﺮرﺳﻲ ﻋﻮاﻣﻞ ﺣﺪت ﺑﺎروش Multiplex PCR ﺟﻬﺖ ﺗﺸﺨﻴﺺ ژن ﻫﺎي اﺧﺘﺼﺎﺻﻲ اﺳﺘﻔﺎده ﺷﺪ. ﻫﻤﭽﻨﻴﻦ آزﻣﺎﻳﺶ دﻳﺴﻚ دﻳﻔﻴﻮژن و E-test ﺑﺮ اﺳﺎس روش CLSI ﺑﺎ آﻧﺘﻲ ﺑﻴﻮﺗﻴﻜﻬﺎي ﻣﺨﺘﻠﻒ اﻧﺠﺎم ﮔﺮدﻳﺪ. ﻳﺎﻓﺘﻪﻫﺎ: ﻧﺘﺎﻳﺞ آﻧﺘﻲﺑﻴﻮﮔﺮام ﺟﺪاﻳﻪﻫﺎي اﻧﺴﺎﻧﻲ ﺳﻮدوﻣﻮﻧﺎس آﺋﺰوژﻳﻨـﻮزا ﻧﺸـﺎن داد ﻛـﻪ ﺑﻴﺸـﺘﺮﻳﻦ ﻣﻴـﺰان ﻣﻘﺎوﻣـﺖ ﻧﺴـﺒﺖ ﺑـﻪ آﻧﺘﻲﺑﻴﻮﺗﻴﻚ ﻫﺎي آﻣﭙﻲﺳﻴﻠﻴﻦ و ﺳﻔﭙﻴﻢ )100 %(، و ﺑﻴﺸﺘﺮﻳﻦ ﻣﻴﺰان ﻣﻘﺎوﻣﺖ در ﺟﺪاﻳﻪﻫﺎي داﻣﻲ ﻧﺴﺒﺖ ﺑﻪ آﻣﭙﻲﺳـﻴﻠﻴﻦ 100% وﺟﻮد دارد. ﺷﻴﻮع ژنoprI در 100 درﺻﺪ ﺟﺪاﻳﻪ ﻫﺎي اﻧﺴﺎﻧﻲ ﺷﻨﺎﺳﺎﻳﻲ و ﺗﻤﺎم ﻧﻤﻮﻧـﻪ ﻫـﺎ از ﻧﻈـﺮ وﺟـﻮد ژن alg-D ﻣﻨﻔـﻲ ﺑﻮدﻧﺪ. ﺑﺮﺧﻼف ﻧﻤﻮﻧﻪ ﻫﺎي اﻧﺴﺎﻧﻲ، ﺗﻤﺎﻣﻲ ﻧﻤﻮﻧﻪ ﻫﺎي داﻣﻲ از ﻧﻈﺮ وﺟﻮد اﮔﺰوآﻧـﺰﻳﻢ S ﻣﻨﻔـﻲ ﺑـﻮده و ﻓﻘـﻂ اﮔﺰوآﻧـﺰﻳﻢ Y در 15ﺟﺪاﻳﻪ ﺷﻨﺎﺳﺎﻳﻲ ﺷﺪ. در ﺟﺪاﻳﻪ ﻫﺎي اﻧﺴﺎﻧﻲ اﮔﺰوآﻧﺰﻳﻢ T از ﺑﻴﺸﺘﺮﻳﻦ ﻓﺮاواﻧﻲ ﺑﺮﺧﻮردار ﺑﻮد. ﺑﻪ ﮔﻮﻧـﻪ اي ﻛـﻪ 100% ﻧﻤﻮﻧﻪ ﻫﺎ از ﻧﻈﺮ اﻳﻦ اﮔﺰوآﻧﺰﻳﻢ ﻣﺜﺒﺖ ﺑﻮدﻧﺪ. ﺑﺤﺚ: ﺷﻴﻮع ﺑﺎﻻي ژﻧﻬﺎي ﺣﺪت در ﺑﻴﻦ اﻳﺰوﻟﻪ ﻫﺎي اﻧﺴﺎﻧﻲ و داﻣﻲ و ﺗﻮﺟﻪ ﺑﻪ ﻋﻤﻠﻜﺮد اﻳﻦ ژنﻫﺎ، ﺷﻨﺎﺳﺎﻳﻲ ﺗﺎﺛﻴﺮ ﺑﺮ ﺑﺎﻓﺖ ﻫﺎي داﻣﻲ و اﻧﺴﺎﻧﻲ را ﺿﺮوري ﻣﻲﺳﺎزد. ﺑﺎ ﺗﻮﺟﻪ ﺑﻪ ﺑﺎﻻ ﺑﻮدن ﻣﻴﺰان ﻣﻘﺎوﻣﺖ آﻧﺘﻲﺑﻴﻮﺗﻴﻜﻲ در ﺳﻮﻳﻪ ﻫﺎي داﻣﻲ و اﻧﺴـﺎﻧﻲ، ﻣﺼـﺮف ﻣﻨﺎﺳﺐ و ﺑﻪ اﻧﺪازه آﻧﺘﻲ ﺑﻴﻮﺗﻴﻚ ﻫﺎ ﻣﻲﺑﺎﻳﺴﺖ رﻋﺎﻳﺖ ﮔﺮدد.

Background and Aims: Pseudomonas aeruginosa is the most important cause of various nosocomial infections and mastitis in dairy cattle and the development of antibiotic resistance. The aim of this study was to determine the antibiotic resistance of Pseudomonas aeruginosa and the presence of virulence genes in human and animal samples. Materials and Methods: In this study, 102 human and animal strains of Pseudomonas aeruginosa were studied. Multiplex PCR for virulence factors was used to detect specific genes. Disk diffusion and E-test were performed according to CLSI method with different antibiotics. Results: The results of antibiogram of human isolates of Pseudomonas aeruginosa showed the highest resistance to ampicillin and cefepime antibiotics (100%), and the highest resistance in animal isolates to ampicillin (100%). The prevalence of oprI gene was detected in 100% of human isolates and all samples were negative for alg-D gene. Unlike human samples, all animal samples were negative for exoenzyme S and only exoenzyme Y was detected in 15 isolates (25%). Exoenzyme T was the most abundant in human isolates. 100% of the samples were positive for this exoenzyme. Conclusion: High prevalence of virulence genes between human and animal isolates and attention to the function of these genes makes it necessary to identify the effect on animal and human tissues. Due to the high level of antibiotic resistance in animal and human strains, proper use and size of antibiotics should be observed.