title: A Novel Loop-Mediated Isothermal Amplification Assay for Serogroup Identification of Neisseria meningitidis in Cerebrospinal Fluid.
abstract: We have developed a novel Neisseria meningitidis serogroup-specific loop-mediated isothermal amplification (LAMP) assay for six of the most common meningococcal serogroups (A, B, C, W, X, and Y). The assay was evaluated using a set of 31 meningococcal LAMP assay positive cerebrospinal fluid (CSF) specimens from 1574 children with suspected meningitis identified in prospective surveillance between 1998 and 2002 in Vietnam, China, and Korea. Primer specificity was validated using 15 N. meningitidis strains (including serogroups A, B, C, E, W, X, Y, and Z) and 19 non-N. meningitidis species. The N. meningitidis serogroup LAMP detected down to ten copies and 100 colony-forming units per reaction. Twenty-nine CSF had N. meningitidis serogroup identified by LAMP compared with two CSF in which N. meningitidis serogroup was identified by culture and multi-locus sequence typing. This is the first report of a serogroup-specific identification assay for N. meningitidis using the LAMP method. Our results suggest that this assay will be a rapid, sensitive, and uniquely serogroup-specific assay with potential for application in clinical laboratories and public health surveillance systems.
authors.affiliation: Department of Pharmacy, College of Pharmacy, Hanyang UniversityAnsan, South Korea; Institute of Pharmacological Research, Hanyang UniversityAnsan, South Korea.
authors.forename: DoKyung
authors.initials: D
authors.lastname: Lee
authors.affiliation: Department of Pharmacy, College of Pharmacy, Hanyang UniversityAnsan, South Korea; Institute of Pharmacological Research, Hanyang UniversityAnsan, South Korea.
authors.forename: Eun Jin
authors.initials: EJ
authors.lastname: Kim
authors.affiliation: Department of Pharmacy Practice, Eugene Applebaum College of Pharmacy & Health Sciences, Wayne State University Detroit, MI, USA.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Kilgore
authors.affiliation: Department of Bacteriology I, National Institute of Infectious Diseases Tokyo, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Takahashi
authors.affiliation: Department of Bacteriology I, National Institute of Infectious Diseases Tokyo, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Ohnishi
authors.affiliation: Kaneka, Co., Ltd., Osaka Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Tomono
authors.affiliation: Kaneka, Co., Ltd., Osaka Japan.
authors.forename: Shigehiko
authors.initials: S
authors.lastname: Miyamoto
authors.affiliation: Nihon University School of DentistryTokyo, Japan; Dental Research Center, Nihon University School of DentistryTokyo, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Omagari
authors.affiliation: Department of Pharmacy, College of Pharmacy, Hanyang UniversityAnsan, South Korea; Institute of Pharmacological Research, Hanyang UniversityAnsan, South Korea.
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Kim
authors.affiliation: Nihon University School of DentistryTokyo, Japan; Dental Research Center, Nihon University School of DentistryTokyo, Japan.
authors.forename: Mitsuko
authors.initials: M
authors.lastname: Seki
keywords: Neisseria meningitidis; cerebrospinal fluid; children; loop-mediated isothermal amplification; meningitis; serogroup identification