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.
Highlights
Neisseria meningitidis is a gram-negative β-proteobacterium and member of the bacterial family Neisseriaceae (Stephens et al, 2007)
We report the development of a loop-mediated isothermal amplification (LAMP)-based N. meningitidis serogroup identification assay and the evaluation of this assay’s ability to detect specific N. meningitidis serogroups in cerebrospinal fluid (CSF)
We evaluated the serogroup identification LAMP assay using the set of 31 meningococcal LAMP (Nm-LAMP) positive CSF
Summary
Neisseria meningitidis is a gram-negative β-proteobacterium and member of the bacterial family Neisseriaceae (Stephens et al, 2007). The meningococcal polysaccharide capsule and outer membrane proteins have been identified as N. meningitidis virulence factors (Stephens, 2009). N. meningitidis is principally subdivided into 12 serogroups based on the capsular polysaccharide type (A, B, C, E, H, I, K, L, W, X, Y, and Z; Harrison et al, 2013). The majority of invasive meningococcal disease has been attributable to six serogroups designated A, B, C, W, X, and Y (Stephens et al, 2007; Stephens, 2009; Verma and Khanna, 2012).
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