Detsika Maria G, Corkill John E, Magalhães Marcelo, Glendinning Kerry J, Hart C Anthony, Winstanley Craig
Department of Medical Microbiology and Genitourinary Medicine, University of Liverpool, Liverpool L69 3GA, United Kingdom.
J Clin Microbiol. 2003 Sep;41(9):4148-53. doi: 10.1128/JCM.41.9.4148-4153.2003.
PCR tests were used to assign genomovar status to 39 non-cystic fibrosis (non-CF) and 11 CF Burkholderia cepacia complex isolates from patients in hospitals in Recife, Brazil. Non-CF isolates were assigned to genomovar IIIA (71.8%), genomovar I (15.4%), B. vietnamiensis (7.7%), and B. multivorans (5.1%). CF isolates were assigned to genomovar IIIA (18.2%), B. vietnamiensis (18.2%), and genomovar I (9.1%). Six CF isolates sharing recA PCR-restriction fragment length polymorphism (RFLP) and randomly amplified polymorphic DNA (RAPD) patterns could not be assigned to a genomovar. 16S rDNA sequence obtained from these isolates indicated a closest relationship to B. anthina, but the recA sequence was equally divergent from several genomovars. PCR screening indicated the presence of cblA in only two isolates, whereas the B. cepacia epidemic strain marker was found in 22 of 28 genomovar IIIA isolates. A type III secretion gene was detected in all but genomovar I isolates. RAPD and PCR-RFLP assays, targeting both recA and fliC, indicated a large amount of genetic variability among the isolates, with many novel patterns being observed. Nine genomovar IIIA isolates from different non-CF patients and clinical sources had identical genotypes, indicating the presence of a common clone.
采用聚合酶链反应(PCR)检测法,对来自巴西累西腓市医院患者的39株非囊性纤维化(非CF)伯克霍尔德菌复合体菌株和11株CF伯克霍尔德菌复合体菌株进行基因分型。非CF菌株被分为基因变种IIIA(71.8%)、基因变种I(15.4%)、越南伯克霍尔德菌(7.7%)和多食伯克霍尔德菌(5.1%)。CF菌株被分为基因变种IIIA(18.2%)、越南伯克霍尔德菌(18.2%)和基因变种I(9.1%)。6株CF菌株因共享recA聚合酶链反应-限制性片段长度多态性(RFLP)和随机扩增多态性DNA(RAPD)图谱而无法进行基因分型。从这些菌株获得的16S核糖体DNA序列显示与炭疽伯克霍尔德菌关系最为密切,但recA序列与多个基因变种的差异程度相同。PCR筛查表明仅在2株菌株中存在cblA,而在28株基因变种IIIA菌株中的22株中发现了洋葱伯克霍尔德菌流行株标记。除基因变种I菌株外,在所有菌株中均检测到III型分泌基因。针对recA和fliC的RAPD和PCR-RFLP分析表明,这些菌株之间存在大量遗传变异,观察到许多新的图谱。来自不同非CF患者和临床来源的9株基因变种IIIA菌株具有相同的基因型,表明存在一个共同克隆。