CAJ | 학술논문

目的探讨体外人工诱导的耐头孢曲松淋球菌的分子耐药机制.方法在成功诱导淋球菌标准菌株ATCC49226和临床菌株ZSSY00205对头孢曲松耐药的基础上,分别对标准菌株和临床菌株进行诱导后耐药株对诱导前敏感株的抑制性消减杂交,构建差异基因文库.从文库中随机挑取192个差异基因作为探针点样于基因芯片,用分别标记Cy3、Cy5的敏感株、耐药株基因组DNA的RsaI酶切片段同时与芯片杂交,根据芯片扫描图选取差异荧光探针对应的基因进行测序和Blast分析.结果分别构建淋球菌标准菌株ATCC49226和临床菌株ZSSY00205的诱导后耐药株DNA特异性的消减文库,并分别获得高分辨的基因芯片扫描图,发现两组菌株间有共同的耐药相关基因mtrR、mtrC、gyrB、rpsJ、PJD1.结论淋球菌对头孢曲松耐药与mtrR、mtrC、gyrB、rpsJ等基因有关,很可能通过外排泵活性增强的途径介导耐药.伴随淋球菌对头孢曲松产生耐药的同时,可能引起对青霉素、四环素、红霉素、喹诺酮等多种抗菌药物产生耐药,即多重耐药现象.
목적 탐토체외인공유도적내두포곡송림구균적분자내약궤제.방법 재성공유도림구균표준균주ATCC49226화림상균주ZSSY00205대두포곡송내약적기출상,분별대표준균주화림상균주진행유도후내약주대유도전민감주적억제성소감잡교,구건차이기인문고.종문고중수궤도취192개차이기인작위탐침점양우기인심편,용분별표기Cy3、Cy5적민감주、내약주기인조DNA적RsaI매절편단동시여심편잡교,근거심편소묘도선취차이형광탐침대응적기인진행측서화Blast분석.결과 분별구건림구균표준균주ATCC49226화림상균주ZSSY00205적유도후내약주DNA특이성적소감문고,병분별획득고분변적기인심편소묘도,발현량조균주간유공동적내약상관기인mtrR、mtrC、gyrB、rpsJ、PJD1.결론 림구균대두포곡송내약여mtrR、mtrC、gyrB、rpsJ등기인유관,흔가능통과외배빙활성증강적도경개도내약.반수림구균대두포곡송산생내약적동시,가능인기대청매소、사배소、홍매소、규낙동등다충항균약물산생내약,즉다중내약현상.
Objective To elucidate the molecular basis for induced resistance of N. gonorrhoeae to ceftriaxone in vitro. Methods The reference strain ATCC49226 and clinical isolate ZSSY00205 of N. gon-orrhoeae were exposed to subinhibitory concentration of ceftriaxone for the induction of resistance. Then,suppression subtractive hybridization was performed with the pre-induction parent strains as drivers and post-induction mutant strains as testers to create a subtractive cDNA library. Following that, a total of 192 clones were randomly selected from the library, and arrayed by spotting onto nylon membranes. Finally, dif-ferentially expressed genes were screened by hybridization with labeled-RsaI restriction fragments from the sensitive and resistant N.gonorrhoeae strains respectively, and analyzed by sequencing and homology research using Blast program. Results A subtractive library for these resistant N.gonorrhoeae strains was generated by SSH technique. Microarray analysis and homology research confirmed 5 genes related to ceftriaxone resistance, i.e. mtrR, mtrC, gyrB, rpsJ and PJD1. Conclusions The induced resistance of N. gonorrhoeae to ceftriaxone may be associated with mtrR, mtrC, gyrB, rpsJ and PJD1 genes which probably mediate the resistance by enhancing the activity of efflux pump system.