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Loop-Mediated Isothermal Amplification Methods For Diagnosis Of Bacterial Meningitis, Mitsuko Seki, Paul E. Kilgore, Eun Jin Kim, Makoto Ohnishi, Satoshi Hayakawa, Dong Wook Kim

Department of Pharmacy Practice

The rapid, accurate, and efficient identification of an infectious disease is critical to ensure timely clinical treatment and prevention in public health settings. In 2015, meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis was responsible for 379,200 (range: 322,700–444,700) deaths. Clinical features alone cannot determine whether bacterial meningitis is present; an analysis of cerebrospinal fluid (CSF) is essential. Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification method offering an alternative to polymerase chain reaction (PCR). LAMP-based assays for detection of three leading bacteria in CSF for diagnosis of meningitis have been established. The typing assays …

A Novel Loop-Mediated Isothermal Amplification Assay For Serogroup Identification Of Neisseria Meningitidis In Cerebrospinal Fluid, Dokyung Lee, Eun Jin Kim, Paul E. Kilgore, Hideyuki Takahashi, Jun Tomono, Shigehiko Miyamoto, Daisuke Omagari, Dong Wook Kim, Mitsuko Seki

Department of Pharmacy Practice

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 …

An Expanded Age Range For Meningococcal Meningitis: Molecular Diagnostic Evidence From Population-Based Surveillance In Asia, Soon Kim, Dong Kim, Bai Dong, Jung Kim, Dang Anh, Paul E. Kilgore

Wayne State University Associated BioMed Central Scholarship

Abstract

Background

To understand epidemiologic patterns of meningococcal disease in Asia, we performed a retrospective molecular analysis of cerebrospinal fluid (CSF) specimens collected in prospective surveillance among children aged < 5 years of age in China, South Korea, and Vietnam.

Methods

A total of 295 isolates and 2,302 CSFs were tested by a meningococcal species- and serogroup-specific polymerase chain reaction (PCR) assay targeting the Neisseria meningitidis (Nm) ctrA gene. Multi-locus sequence typing (MLST) was performed in Nm gene amplification analysis and incidence rates for meningococcal meningitis were estimated.

Results

Among 295 isolates tested, 10 specimens from Vietnam were confirmed as serogroup B and all were Sequence Type (ST) 1576 by MLST. …