TY - BOOK AU - Amr Elsayed Ibrahim Ahmed Elghopashy, AU - Yasser El Mohammady Ragab AU - Aymen Samir Yassin AU - Mohamed Abd El-Gawad El-Sayed Ahmed TI - Molecular study on carbapenem resistant klebsiella pneumoniae isolated from egyptian hospitals U1 - 616.01 PY - 2025/// KW - Microbiology KW - الميكوبيولوجيا الطبية KW - Carbapenem KW - resistant Klebsiella pneumoniae (CR-KP) KW - Carbapenems KW - MDR KW - Carbapenemases KW - blaOXA-48 KW - blaNDM-1 N1 - Thesis (M.Sc)-Cairo University, 2025; Bibliography: pages 119-146.; Issues also as CD N2 - Carbapenem-resistant Klebsiella pneumoniae (CR-KP) infections are considered a global critical concern, causing a serious threat to public health. CR-KP strains have been widely disseminated worldwide in recent years, affecting countries with varying income levels, including low-, middle-, and upper-income countries. The incidence of CR-KP infections is tremendously increasing in developing countries, such as Egypt, posing a significant threat to public health. Ninety-four clinically significant K. pneumoniae isolates were collected from two tertiary care Egyptian hospitals: Al-Kasr Al-Ainy hospital, Cairo, Egypt and Memorial Souad Kafafi University hospital, Giza, Egypt over one year. All collected isolates were subcultured, and their identification as K. pneumoniae ssp. pneumoniae was confirmed via the automated VITEK® 2 system and the MALDI-TOF MS. Antimicrobial susceptibility testing (AST) of all tested K. pneumoniae isolates was examined using the Kirby-Bauer disk diffusion method and MICs were determined by the automated VITEK® 2 system, then confirmed for imipenem, meropenem, and colistin via the routine broth microdilution method. Subsequently, phenotypic detection of carbapenemases, as well as genotypic detection of major carbapenemase-encoding genes were carried out. Based on the automated VITEK® 2 system's AST results, all clinical K. pneumoniae isolates were multidrug-resistant (MDR). The highest detected resistance patterns were toward β- lactam antibiotics, followed by quinolones and aminoglycosides, where the prevalence of carbapenems resistance was too high, with 94.68% of isolates being resistant to imipenem with MICs >16 µg/ml. Phenotypic assays showed that 98.9% of CR-KP isolates produced carbapenemase(s), while PCR technique revealed that 97.9% of tested isolates harbored at least one carbapenemase-encoding gene. BlastN analysis of the sequenced PCR products against GenBank's non-redundant genome sequences showed that the blaOXA-48 gene was the most predominant carbapenemase-encoding genotype among our isolates, found in 89/94 of the tested isolates (94.7%), followed by blaNDM-1 (65/94, 69.1%) and blaKPC-2 (5/94, 5.3%). Additionally, the co-existence of both blaOXA-48 and blaNDM-1 carbapenemase genes in certain isolates (67%) was also observed. The high prevalence of carbapenem resistance and the terrifying spread of their encoding genes among K. pneumoniae isolates in Egypt are alarming, leading to limitations in the available therapeutic options for the management of these serious infections. Carbapenemases of the blaOXA- 48 and blaNDM-1 genotypes are widespread in Egyptian healthcare settings, causing the current increase in carbapenem-resistant bacteria; ﺗﻌﺘﺒﺮ ﻋﺪوى اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ اﻟﻤﻘﺎوﻣﺔ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت ﻣﺼﺪر ﻗﻠﻖ ﻋﺎﻟﻤﻲ ﺑﺎﻟﻎ ﯾﮭﺪد اﻟﺼﺤﺔ اﻟﻌﺎﻣﺔ ﺑﺸﻜﻞ ﺧﻄﯿﺮ. وﻗﺪ اﻧﺘﺸﺮت ھﺬه اﻟﺴﻼﻻت ﻣﻦ اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ اﻟﻤﻘﺎوﻣﺔ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت ﺑﺸﻜﻞ واﺳﻊ ﻓﻲ ﺟﻤﯿﻊ أﻧﺤﺎء اﻟﻌﺎﻟﻢ ﻓﻲ اﻟﺴﻨﻮات اﻷﺧﯿﺮة، ﺣﯿﺚ أﺛﺮت ﻋﻠﻰ اﻟﺒﻠﺪان ذات ﻣﺴﺘﻮﯾﺎت اﻟﺪﺧﻞ اﻟﻤﺨﺘﻠﻔﺔ، ﺑﻤﺎ ﻓﻲ ذﻟﻚ اﻟﺒﻠﺪان ذات اﻟﺪﺧﻞ اﻟﻤﻨﺨﻔﺾ واﻟﻤﺘﻮﺳﻂ واﻟﻤﺮﺗﻔﻊ. ﻓﯿﻤﺎ ﺗﺸﮭﺪ اﻟﺒﻠﺪان اﻟﻨﺎﻣﯿﺔ ﻣﺜﻞ ﻣﺼﺮ زﯾﺎدة ھﺎﺋﻠﺔ ﻓﻲ ﻣﻌﺪل اﻧﺘﺸﺎر ﺗﻠﻚ اﻟﻌﺪوى ﻣﻤﺎ ﯾﺸﻜﻞ ﺧﻄﺮًا ﻛﺒﯿﺮًا ﻋﻠﻰ اﻟﺼﺤﺔ اﻟﻌﺎﻣﺔ. ﺗﻢ ﺟﻤﻊ أرﺑﻌﺔ وﺗﺴﻌﻮن ﻋﯿﻨﺔ ﻣﻦ اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ ذات اﻷھﻤﯿﺔ اﻟﺴﺮﯾﺮﯾﺔ ﻣﻦ ﻣﺴﺘﺸﻔﯿﯿﻦ ﻣﺼﺮﯾﺘﯿﻦ ﻋﻠﻰ ﻣﺪار ﻋﺎم واﺣﺪ. ﺗﻢ إﻋﺎدة زراﻋﺔ ﺟﻤﯿﻊ اﻟﻌﯿﻨﺎت اﻟﻤﺠﻤﻌﺔ وﺗﻢ ﺗﺄﻛﯿﺪ ﺗﻌﺮﯾﻔﮭﺎ ﻋﻠﻰ أﻧﮭﺎ ﻋﯿﻨﺎت ﻛلبسيله رﺋﻮﯾﺔ ﺑﺎﺳﺘﺨﺪام ﻧﻈﺎم 2 VITEK® اﻵﻟﻲ وﻣﻦ ﺧﻼل MS .MALDI-TOF ﺗﻢ ﻓﺤﺺ ﺣﺴﺎﺳﯿﺔ ﺟﻤﯿﻊ ﻋﺰﻻت اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ ﺗﺠﺎه اﻟﻤﻀﺎدات اﻟﺤﯿﻮﯾﺔ اﻟﻤﺨﺘﻠﻔﺔ ﺑﺎﺳﺘﺨﺪام طﺮﯾﻘﺔ diffusion disk Kirby-Bauer، وﺗﻢ ﺗﺤﺪﯾﺪ ﺗﺮاﻛﯿﺰ اﻟﺤﺪ اﻷدﻧﻰ اﻟﻤﺜﺒﻂ ﻟﻠﻨﻤﻮ ﻟﻠﻤﻀﺎدات اﻟﻤﯿﻜﺮوﺑﯿﺔ ﺑﺎﺳﺘﺨﺪام ﻧﻈﺎم 2 VITEK® اﻵﻟﻲ، وﺗﻢ ﺗﺄﻛﯿﺪھﺎ ﻟﻺﯾﻤﯿﺒﯿﻨﯿﻢ واﻟﻤﯿﺮوﺑﯿﻨﯿﻢ واﻟﻜﻮﻟﯿﺴﺘﯿﻦ ﺑﻮاﺳﻄﺔ طﺮﯾﻘﺔmicrodilution .broth ﺛﻢ ﺑﻌﺪ ذﻟﻚ ﺗﻢ إﺟﺮاء اﻟﻜﺸﻒ اﻟﻈﺎھﺮي ﻋﻦ إﻧﺘﺎج اﻟﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎزات وﻛﺬﻟﻚ اﻟﻜﺸﻒ اﻟﺠﯿﻨﻲ ﻟﻠﺠﯿﻨﺎت اﻟﺮﺋﯿﺴﯿﺔ اﻟﻤﻨﺘﺠﮫ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎزات. اﺳﺘﻨﺎداً إﻟﻰ ﻧﺘﺎﺋﺞ اﺧﺘﺒﺎر اﻟﺤﺴﺎﺳﯿﺔ ﻟﻠﻤﻀﺎدات اﻟﻤﯿﻜﺮوﺑﯿﺔ ﺑﺎﺳﺘﺨﺪام ﻧﻈﺎم 2 VITEK® اﻵﻟﻲ، ﻛﺎﻧﺖ ﺟﻤﯿﻊ ﻋﺰﻻت اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ اﻟﺴﺮﯾﺮﯾﺔ ﻣﻘﺎوﻣﺔ ﻣﺘﻌﺪدة ﻟﻠﻤﻀﺎدات اﻟﺤﯿﻮﯾﺔ .(MDR) ﻛﺎﻧﺖ أﻧﻤﺎط اﻟﻤﻘﺎوﻣﺔ اﻷﻋﻠﻰ اﻟﻤﻜﺘﺸﻔﺔ ﺗﺠﺎه اﻟﻤﻀﺎدات اﻟﺤﯿﻮﯾﺔ ﻣﻦ ﻧﻮع ﺑﯿﺘﺎ-ﻻﻛﺘﺎم، ﺗﻠﯿﮭﺎ اﻟﻔﻠﻮروﻛﯿﻨﻮﻟﻮﻧﺎت واﻷوﻣﯿﻨﻮﺟﻠﯿﻜﻮﺳﯿﺪات، ﻓﯿﻤﺎ ﻛﺎﻧﺖ ﻧﺴﺒﺔ اﻟﻤﻘﺎوﻣﺔ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت ﻋﺎﻟﯿﺔ ﻟﻠﻐﺎﯾﺔ، ﺣﯿﺚ ﻛﺎﻧﺖ ٦۸٫۹٤٪ ﻣﻦ اﻟﻌﺰﻻت ﻣﻘﺎوﻣﺔ ﻟﻺﯾﻤﯿﺒﯿﻨﯿﻢ ﻣﻊ ﺣﺪ أدﻧﻰ ﻣﺜﺒﻂ ﻟﻠﻨﻤﻮ أﻛﺒﺮ ﻣﻦ ۱٦ ﻣﯿﻜﺮوﻏﺮام/ﻣﻞ. وأظﮭﺮت اﻟﻔﺤﻮﺻﺎت اﻟﻈﺎھﺮﯾﺔ أن ۹٫۹۸٪ ﻣﻦ ﻋﺰﻻت اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ اﻟﻤﻘﺎوﻣﺔ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت ﺗﻨﺘﺞ ﻛﺎرﺑﺎﺑﯿﻨﯿﻤﯿﺰ، ﺑﯿﻨﻤﺎ ﻛﺸﻔﺖ ﺗﻘﻨﯿﺔ ﺗﻔﺎﻋﻞ ﺗﺴﻠﺴﻞ اﻟﺒﻠﻤﺮة أن ۹٫۹۷٪ ﻣﻦ اﻟﻌﺰﻻت اﻟﻤﺨﺘﺒﺮة ﺗﺤﺘﻮي ﻋﻠﻰ ﺟﯿﻦ واﺣﺪ ﻋﻠﻰ اﻷﻗﻞ ﻣﻨﺘﺞ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﯿﺰ. ﻛﻤﺎ أظﮭﺮ ﺗﺤﻠﯿﻞ BlastN ﻟﻤﻨﺘﺠﺎت ﺗﻔﺎﻋﻞ ﺗﺴﻠﺴﻞ اﻟﺒﻠﻤﺮة اﻟﻤﺘﺴﻠﺴﻠﺔ ﻣﻘﺎﺑﻞ ﺗﺴﻠﺴﻼت اﻟﺠﯿﻨﻮم ﻏﯿﺮ اﻟﻤﺘﻜﺮرة ﻓﻲ ﺑﻨﻚ اﻟﺠﯿﻨﺎت أن ﺟﯿﻦ blaOXA-48 ﻛﺎن اﻟﻨﻤﻂ اﻟﺠﯿﻨﻲ اﻷﻛﺜﺮ اﻧﺘﺸﺎرًا ﺑﯿﻦ اﻟﻌﺰﻻت ﻟﺪﯾﻨﺎ، ﺣﯿﺚ وﺟﺪ ﻓﻲ ۸۹ ﻣﻦ أﺻﻞ ۹٤ ﻋﯿﻨﺔ ۷)٫۹٤٪(، ﯾﻠﯿﮫ blaNDM-1 ۹٤/٦٥)، ۱٫٦۹٪( و blaKPC-2 ۹٤/٥)، ۳٫٥٪.( ﺑﺎﻹﺿﺎﻓﺔ إﻟﻰ ذﻟﻚ، ﺗﻢ ﻣﻼﺣﻈﺔ وﺟﻮد ﺟﯿﻨﻲ blaOXA-48 وblaNDM-1 ﻓﻲ ﻋﺰﻻت ﻣﻌﯿﻨﺔ بنسبة .٪٦۷ إن اﻻرﺗﻔﺎع اﻟﻜﺒﯿﺮ ﻓﻲ ﻣﻘﺎوﻣﺔ اﻟﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت واﻻﻧﺘﺸﺎر اﻟﻤﺮوع ﻟﻠﺠﯿﻨﺎت اﻟﻤﻨﺘﺠﺔ ﻟﮭﺎ ﺑﯿﻦ ﻋﺰﻻت اﻟﻜﻠﺒﺴﯿﻠﮫ اﻟﺮﺋﻮﯾﺔ ﻓﻲ ﻣﺼﺮ ﯾﻌﺪ أﻣﺮًا ﻣﺜﯿﺮًا ﻟﻠﻘﻠﻖ، ﻣﻤﺎ ﯾﺆدي إﻟﻰ ﻣﺤﺪودﯾﺔ اﻟﺨﯿﺎرات اﻟﻌﻼﺟﯿﺔ اﻟﻤﺘﺎﺣﺔ ﻟﻠﺘﻌﺎﻣﻞ ﻣﻊ ھﺬه اﻟﻌﺪوى اﻟﺨﻄﯿﺮة. ﺗﻌﺘﺒﺮ ﻛﺎرﺑﺎﺑﯿﻨﯿﻤﯿﺰات ﻣﻦ اﻟﻨﻮع اﻟﺠﯿﻨﻲ blaOXA-48 و blaNDM-1 ﻣﻨﺘﺸﺮة ﺑﺸﻜﻞ واﺳﻊ ﻓﻲ اﻟﺒﯿﺌﺎت اﻟﺼﺤﯿﺔ اﻟﻤﺼﺮﯾﺔ، ﻣﻤﺎ ﯾﺘﺴﺒﺐ ﻓﻲ اﻟﺰﯾﺎدة اﻟﺤﺎﻟﯿﺔ ﻓﻲ اﻟﺒﻜﺘﯿﺮﯾﺎ اﻟﻤﻘﺎوﻣﺔ ﻟﻠﻜﺎرﺑﺎﺑﯿﻨﯿﻤﺎت ER -