【摘要】：目的： 建立脈沖場凝膠電泳(Pulsed-Field Gel Electrophoresis, PFGE)基因分型方法,對天津市南開醫(yī)院和天津市兒童醫(yī)院臨床分離的銅綠假單胞菌(Pseudomonas aeruginosa, PA)進行基因分型,進而對PA進行流行病學分析。 方法： (1)臨床送檢標本按《全國臨床檢驗操作規(guī)程》鑒定為PA,質(zhì)控菌株為PAATCC27853。 (2)采用K-B紙片擴散法對PA進行藥敏試驗,測試其對不同抗菌藥物的敏感性,藥敏結(jié)果按美國CLSI標準判讀。南開醫(yī)院PA的藥敏試驗檢測14種抗菌藥物,兒童醫(yī)院PA的藥敏試驗檢測15種抗菌藥物。 (3)對兩家醫(yī)院臨床分離得到的128株PA進行PFGE基因分型和流行病學分析。 (4)對臨床分離出PA宿主的臨床病例進行回顧性分析,判斷菌株分離時段是否處在感染狀態(tài)。 結(jié)果： (1)感染和定植菌株的判斷：來自于南開醫(yī)院的72株PA中,有57株(79%)菌可能為感染菌株；而來自于兒童醫(yī)院的56株PA中,有39株(70%)可能為感染菌株。兩家醫(yī)院的定植菌株均來自于呼吸道。 (2)菌株的臨床來源和分布：兩家醫(yī)院的PA菌株主要分離于痰標本,南開醫(yī)院有51株,占71%；兒童醫(yī)院為51株,占90%；其他菌株分離于膽汁、腹腔液、膿性分泌物、血液和尿液標本。南開醫(yī)院的PA菌株主要來自于中西結(jié)合腦病科和呼吸科,而兒童醫(yī)院菌株主要分離于呼吸科和內(nèi)分泌科。 (3)菌株的藥物敏感性：南開醫(yī)院的PA菌株對14種抗菌藥物的耐藥率在30%-70%之間；但兒童醫(yī)院的PA菌株對抗生素的敏感性與南開醫(yī)院菌株有所不同,耐藥率在0%-100%之間,兒童醫(yī)院菌株均對氨芐西林耐藥,對丁胺卡那霉素、環(huán)丙沙星敏感。 (4) PFGE分型：南開醫(yī)院的72株PA被分為48個PFGE型,Ⅰ型5株；Ⅱ和Ⅲ型各4株；有14種PFGE譜各由2株菌共有,其余31株菌各表現(xiàn)為獨特的PFGE譜。兒童醫(yī)院56株PA被分為18個型別,A型26株；B型12株；C型3株：其余15株各成獨特的PFGE譜。兩家醫(yī)院PA菌株中未見相同或相近的PFGE譜菌株。 (5)兒童醫(yī)院PFGE優(yōu)勢型和耐藥性的關系：兒童醫(yī)院的A和B型菌株幾乎均對4種抗菌藥物耐藥和3種抗菌藥物敏感。但對其余8種藥物兩型表現(xiàn)了不同的耐藥率,其中4種藥物耐藥率有明顯差別。 (6)追溯兩家醫(yī)院主要PFGE型別菌株的時間和空間分布：南開醫(yī)院Ⅰ型菌株是同一患者不同時間分離得到的菌株,Ⅱ型菌株和Ⅲ型菌株由不同科室患者在不同時間分離得到；兒童醫(yī)院A型菌株來自于呼吸科的不同患兒,而B型菌株主要來自內(nèi)分泌科的不同患兒。 結(jié)論： (1)新建立的PFGE方法能夠明確地對PA進行基因分型,可以作為有效的技術手段用于分子流行病學的研究和臨床實踐。 (2)不同醫(yī)院的臨床分離得到的PA菌株抗生素耐藥譜不同。 (3)南開醫(yī)院的PA菌株呈散發(fā)感染的特征,而兒童醫(yī)院PA菌株表現(xiàn)為科室間流行趨勢；兩家醫(yī)院PA菌株無相同基因型,故未見院間傳播；PA菌株進化來源具有遺傳學多元性。 (4) PFGE在基因水平上對PA進行分析,可分析PA的遺傳學特征、感染來源、傳播途徑及分布規(guī)律,對感染流行的監(jiān)測提供可靠依據(jù)。
[Abstract]:Purpose: A pulsed-field gel electrosynthesis (PFGE) genotyping method was established to genotyping the Pseudomonas aeruginosa (PA) isolated from Nankai Hospital and Tianjin Children's Hospital. Analysis. Methods: (1) The clinical test specimens were identified as PA, and the quality control strains were PAATCC. 27853. (2) The susceptibility of PA to different antibacterial drugs was tested by the K-B paper diffusion method. The results of drug sensitivity were as follows: The interpretation of the LSI standard. The drug sensitivity test of the PA in the Nankai Hospital has detected 14 kinds of antibacterial drugs, and the drug sensitivity test of the PA in the children's hospital is tested. 15 antibacterial drugs. (3) The PFGE gene was used for 128 strains of PA isolated from the two hospitals. Typing and epidemiological analysis. (4) The clinical cases of the clinical isolation of the PA host were analyzed retrospectively, and the strains were determined. off period Results: (1) There were 57 strains (79%) of the 72 strains of PA from Nankai Hospital, and 57 (79%) of the strains could be infected, and 56 strains of PA from the children's hospital 39 (70%) may be an infected strain. The strains of colonization in the hospital were from the respiratory tract. (2) The clinical origin and distribution of the strains: the PA strains of the two hospitals were mainly isolated from the sputum specimen, 51 of the Nankai Hospital, and 71%; the children's hospital was 51, accounting for 90%; the other strains were isolated from the bile, Peritoneal fluid, purulent secretion, blood and urine specimen. The PA strain of the Nankai Hospital is mainly from the combination of the Chinese and Western combined encephalopathy and the respiratory department, while the children The hospital strains are mainly isolated from the respiratory and endocrinology. (3) The drug sensitivity of the strain: the resistance rate of the PA strain of the Nankai Hospital to the 14 antibacterial drugs is between 30% and 70%; however, the sensitivity of the PA strain of the children's hospital to the antibiotics is similar to that of the Nankai Hospital. The strains were different and the drug resistance rate was between 0% and 100%, and the children's hospital strains were all sensitive to ammonia. Methicillin-resistant, sensitive to amikacin and ciprofloxacin. (4) PFGE: 72 PA in Nankai Hospital were divided into 48 PFGE-type, 5 strains of type I, 4 strains of type 鈪，

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