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Journal of Clinical Microbiology, August 1999, p. 2402-2407, Vol. 37, No. 8
Meningitis and Special Pathogens Branch,
Received 16 February 1999/Returned for modification 30 March
1999/Accepted 20 April 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.
0095-1137/99/$04.00+0
Cleavase Fragment Length Polymorphism Analysis
of Neisseria meningitidis Basic Metabolic
Genes
*
Corresponding author. Mailing address: Meningitidis and
Special Pathogens Branch, Division of Bacterial and Mycotic Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, MS C-02, Atlanta, GA 30333. Phone: (404) 639-4057. Fax:
(404) 639-3123. E-mail: mlt5{at}cdc.gov.
Journal of Clinical Microbiology, August 1999, p. 2402-2407, Vol. 37, No. 8
0095-1137/99/$04.00+0
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