Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members ...Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members of the complex. This is a single blind study aimed to evaluate certain commonly used species-specific genetic markers namely, Intergenic Transcribed Spacer region in 16S rRNA of <em>A. baumannii</em> (Ab-ITS) and <em>gyrB</em>, for identification of ACB members. These molecular targets were first validated on clinical isolates (n = 200) and subsequently on uncultured tracheal aspirates (n = 172). Among the clinical isolates, 183/200 (91.5%) were positive for Ab-ITS. The clinical isolates 17 (17/200) which are failed to amplify in Ab-ITS PCR were subsequently diagnosed by <em>gyrB</em> PCR as <em>A. calcoaceticus</em> (n = 2), <em>A. pitti</em> (n = 6) and <em>A. nosocomialis</em> (n = 9) but not <em>A. baumannii</em>. Among the tracheal aspirates, 62 samples were reported as sterile in Advanced Expert System of VITEK-2, among the remaining 110 samples, 68.1% (75/110) samples contained Ab-ITS target. Twenty-five of the sterile samples (25/62) were found to contain Ab-ITS target sequence. Since, our sample processing method enabled identification of all the species of ACB complex by PCR even in uncultured tracheal aspirates, adaptation of our protocol would enable same day (6 - 8 h) reporting and help the clinician make evidence based therapeutic decision quickly.展开更多
目的分析重症监护病房(intensive care unit,ICU)临床分离的耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的基因型,为临床的防控提供依据.方法收集云南省第二人民医院ICU临床分离的耐碳青霉烯类鲍曼不动杆菌138株,应用聚合酶链反应(polymerase c...目的分析重症监护病房(intensive care unit,ICU)临床分离的耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的基因型,为临床的防控提供依据.方法收集云南省第二人民医院ICU临床分离的耐碳青霉烯类鲍曼不动杆菌138株,应用聚合酶链反应(polymerase chain reaction,PCR)检测OXA-23、OXA-24、OXA-51、ISAba1-oxa-23、ISAba1-oxa-51共五种碳青霉烯酶基因.结果在138株耐碳青霉烯类鲍曼不动杆菌中,OXA-51型的检出率是97.1%;OXA-23型的检出率是81.3%;ISAba1-oxa-23基因的检出率是65.94%;ISAba1-oxa-51基因的检出率是21.0%.OXA-51+OXA-23+ISAba1-oxa-23型碳青霉烯酶基因表达模式的检出率为44.7%;其次是OXA-51+ISAba1-oxa-51+OXA-23+ISAba1-oxa-23型表达模式,检出率为21.0%.结论OXA-51+OXA-23+ISAba1-oxa-23型碳青霉烯酶基因表达模式是云南省第二人民医院重症监护病房耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的主要表达模式;产OXA-23型碳青霉烯酶是鲍曼不动杆菌对碳青霉烯类抗生素耐药的主要原因.展开更多
文摘Currently, available phenotyping and commercial methods report <em>A. baumannii</em> only as <em>Acinetobacter calcoaceticus-baumannii</em> complex (ACB) and do not identify individual members of the complex. This is a single blind study aimed to evaluate certain commonly used species-specific genetic markers namely, Intergenic Transcribed Spacer region in 16S rRNA of <em>A. baumannii</em> (Ab-ITS) and <em>gyrB</em>, for identification of ACB members. These molecular targets were first validated on clinical isolates (n = 200) and subsequently on uncultured tracheal aspirates (n = 172). Among the clinical isolates, 183/200 (91.5%) were positive for Ab-ITS. The clinical isolates 17 (17/200) which are failed to amplify in Ab-ITS PCR were subsequently diagnosed by <em>gyrB</em> PCR as <em>A. calcoaceticus</em> (n = 2), <em>A. pitti</em> (n = 6) and <em>A. nosocomialis</em> (n = 9) but not <em>A. baumannii</em>. Among the tracheal aspirates, 62 samples were reported as sterile in Advanced Expert System of VITEK-2, among the remaining 110 samples, 68.1% (75/110) samples contained Ab-ITS target. Twenty-five of the sterile samples (25/62) were found to contain Ab-ITS target sequence. Since, our sample processing method enabled identification of all the species of ACB complex by PCR even in uncultured tracheal aspirates, adaptation of our protocol would enable same day (6 - 8 h) reporting and help the clinician make evidence based therapeutic decision quickly.
文摘目的分析重症监护病房(intensive care unit,ICU)临床分离的耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的基因型,为临床的防控提供依据.方法收集云南省第二人民医院ICU临床分离的耐碳青霉烯类鲍曼不动杆菌138株,应用聚合酶链反应(polymerase chain reaction,PCR)检测OXA-23、OXA-24、OXA-51、ISAba1-oxa-23、ISAba1-oxa-51共五种碳青霉烯酶基因.结果在138株耐碳青霉烯类鲍曼不动杆菌中,OXA-51型的检出率是97.1%;OXA-23型的检出率是81.3%;ISAba1-oxa-23基因的检出率是65.94%;ISAba1-oxa-51基因的检出率是21.0%.OXA-51+OXA-23+ISAba1-oxa-23型碳青霉烯酶基因表达模式的检出率为44.7%;其次是OXA-51+ISAba1-oxa-51+OXA-23+ISAba1-oxa-23型表达模式,检出率为21.0%.结论OXA-51+OXA-23+ISAba1-oxa-23型碳青霉烯酶基因表达模式是云南省第二人民医院重症监护病房耐碳青霉烯类鲍曼不动杆菌碳青霉烯酶的主要表达模式;产OXA-23型碳青霉烯酶是鲍曼不动杆菌对碳青霉烯类抗生素耐药的主要原因.