Tondella M L, Reeves M W, Popovic T, Rosenstein N, Holloway B P, Mayer L W
Meningitis and Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
J Clin Microbiol. 1999 Aug;37(8):2402-7. doi: 10.1128/JCM.37.8.2402-2407.1999.
Cleavase fragment length polymorphism (CFLP) is a subtyping system based on the property of the enzyme cleavase to recognize junctions between single- and double-stranded regions of DNA formed after denaturation and cooling. To assess the capacity of CFLP for discriminating Neisseria meningitidis serogroup B strains belonging to the electrophoretic type (ET) 5 (ET-5) complex from other serogroup B strains, 30 serogroup B N. meningitidis isolates were subtyped by CFLP with internal fragments of five housekeeping genes, adk, aspC, carA, dhp, and glnA. Two genes (glnA and carA) which demonstrated a high degree of diversity for the serogroup B isolates were then used to further evaluate the suitability of CFLP for screening 50 serogroup C N. meningitidis outbreak-associated and sporadic-case isolates with a single metabolic gene. The results were compared to those from multilocus enzyme electrophoresis (MEE), the current standard subtyping method. CFLP was able to distinguish the ET-5 complex isolates from other serogroup B isolates as efficiently as MEE. Furthermore, CFLP analysis of a single gene was sufficient to identify and cluster the serogroup C isolates belonging to the ET-37 complex from other, unrelated serogroup C isolates but was not capable of differentiating between the isolates of the major individual ETs of this complex (ET-17 and ET-24) causing most serogroup C meningococcal disease outbreaks in the United States. CFLP based on a single gene with a high degree of diversity but not under selective pressure can be applied to the rapid screening of a large number of isolates related to the recognized epidemic complex ET-5 or ET-37. Additionally, CFLP can be used as an initial screening tool to survey the amount of diversity in genes that might be used to develop a DNA sequence-based subtyping system.
裂解酶片段长度多态性（CFLP）是一种基于裂解酶特性的分型系统，该酶能识别DNA变性和冷却后形成的单链和双链区域之间的连接点。为了评估CFLP区分属于电泳型（ET）5（ET - 5）复合体的B群脑膜炎奈瑟菌菌株与其他B群菌株的能力，用5个管家基因（adk、aspC、carA、dhp和glnA）的内部片段通过CFLP对30株B群脑膜炎奈瑟菌分离株进行分型。然后，使用两个对B群分离株表现出高度多样性的基因（glnA和carA），进一步评估CFLP用于用单个代谢基因筛选50株C群脑膜炎奈瑟菌暴发相关和散发病例分离株的适用性。将结果与当前标准分型方法多位点酶电泳（MEE）的结果进行比较。CFLP能够像MEE一样有效地将ET - 5复合体分离株与其他B群分离株区分开来。此外，对单个基因的CFLP分析足以从其他不相关的C群分离株中识别和聚类属于ET - 37复合体的C群分离株，但无法区分该复合体主要单个ET（ET - 17和ET - 24）的分离株，而这些分离株在美国导致了大多数C群脑膜炎球菌病暴发。基于单个具有高度多样性但不受选择压力影响的基因的CFLP可应用于快速筛选与已识别的流行复合体ET - 5或ET - 37相关的大量分离株。此外，CFLP可用作初始筛选工具，以调查可能用于开发基于DNA序列的分型系统的基因中的多样性程度。
