sodC-Based Real-Time PCR for Detection of Neisseria meningitidis

Thomas, Jennifer D.; Hatcher, Cynthia; Satterfield, Dara A.; Theodore, M. Jordan; Bach, Michelle C.; Linscott, Kristin B.; Zhao, Xin; Wang, Xin; Mair, Raydel; Schmink, Susanna; Arnold, Kathryn E.; Stephens, David S.; Harrison, Lee H.; Hollick, Rosemary; Andrade, Ana Lúcia; Lamaro‐Cardoso, Juliana; Lemos, Ana Paula Silva de; Gritzfeld, Jenna F.; Gordon, Stephen B.; Soysal, Ahmet; Bakır, Mustafa; Sharma, Dolly; Jain, Shabnam; Satola, Sarah W.; Messonnier, Nancy E.; Mayer, Leonard W.

doi:10.1371/journal.pone.0019361

Cited by 81 publications

(48 citation statements)

References 40 publications

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“…Several targets (porA, ctrA, and sodC) have been employed as N. meningitidis-specific diagnostic targets for rt-PCR assays (97,98). In this study, we looked at all three targets by WGS in both of our isolate collections.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Phenotypic and Genotypic Approaches to Capsule Typing of Neisseria meningitidis by Use of Invasive and Carriage Isolate Collections

Mohamed

Rojas

et al. 2016

J Clin Microbiol

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e Neisseria meningitidis serogroup B (MnB) is a leading cause of bacterial meningitis; however, MnB is most commonly associated with asymptomatic carriage in the nasopharyngeal cavity, as opposed to the disease state. Two vaccines are now licensed for the prevention of MnB disease; a possible additional benefit of these vaccines could be to protect against disease indirectly by disrupting nasopharyngeal carriage (e.g., herd protection). To investigate this possibility, accurate diagnostic approaches to characterize MnB carriage isolates are required. In contrast to invasive meningococcal disease (IMD) isolates, which can be readily serogrouped, carriage isolates often lack capsule expression, making standard phenotypic assays unsuitable for strain characterization. Several antibody-based methods were evaluated for their abilities to serogroup isolates and were compared with two genotyping methods (real-time PCR [rt-PCR] and whole-genome sequencing [WGS]) to identify which approach would most accurately ascertain the polysaccharide groups associated with carriage isolates. WGS and rt-PCR were in agreement for 99% of IMD isolates, including those with coding sequences for MnB, MnC, MnW, and MnY, and the phenotypic methods correctly identified serogroups for 69 to 98% of IMD isolates. In contrast, only 47% of carriage isolates were groupable by genotypic methods, due to mutations within the capsule operon; of the isolates identified by genotypic methods, <43% were serogroupable with any of the phenotypic methods tested. These observations highlight the difficulties in the serogrouping and capsular genogrouping of meningococcal carriage isolates. Based on our findings, WGS is the most suitable approach for the characterization of meningococcal carriage isolates. N eisseria meningitidis, a Gram-negative bacterium that causes both epidemic and endemic life-threatening disease, is also an obligate human commensal organism that colonizes the nasopharyngeal mucosa with no or minimal harm to the host, a phenomenon known as carriage (1). Asymptomatic pharyngeal colonization with N. meningitidis in young adults is relatively common, and these asymptomatic carriers represent a potential reservoir for the transmission of pathogenic isolates in the community (2, 3). The rates of asymptomatic carriage in the United States and Europe are highest among adolescents and young adults, peaking at the age of 19 years, and are estimated to be 10% to 35% (4-6). Rates of transmission and carriage are higher in closed and semiclosed populations, such as university students living in dormitories and military recruits housed in barracks (7). Higher rates of carriage are also found among people in close contact with patients with active meningococcal infections (8). For the majority of people, carriage is an immunizing process that results in systemic, serogroup-specific, protective antibody responses (9, 10). Invasive meningococcal disease (IMD) usually occurs shortly after the onset of colonization of a susceptible host, when the bacteria ...

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Section: Discussionmentioning

confidence: 99%

Comparison of Phenotypic and Genotypic Approaches to Capsule Typing of Neisseria meningitidis by Use of Invasive and Carriage Isolate Collections

Mohamed

Rojas

et al. 2016

J Clin Microbiol

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“…A single colony per participant per round was selected for further characterization. At a species level, N. meningitidis was identified using Gram stain, oxidase test (Hardy Diagnostics; Santa Maria, CA), API NH strip (bioMerieux; Durham, NC), and sodC PCR assay [20,21]. Once confirmed as N. meningitidis , serogroup was determined by slide agglutination and capsular genotype by real-time PCR (rt-PCR).…”

Section: Methodsmentioning

confidence: 99%

Meningococcal carriage among a university student population – United States, 2015

Breakwell

Whaley

Khan

et al. 2018

Vaccine

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Objectives Several outbreaks of serogroup B meningococcal disease have occurred among university students in recent years. In the setting of high coverage of the quadrivalent meningococcal conjugate vaccine and prior to widespread use of serogroup B meningococcal vaccines among adolescents, we conducted surveys to characterize the prevalence and molecular characteristics of meningococcal carriage among university students. Methods Two cross-sectional oropharyngeal carriage surveys were conducted among undergraduates at a Rhode Island university. Isolates were characterized using slide agglutination, real-time polymerase chain reaction (rt-PCR), and whole genome sequencing. Adjusted prevalence ratios and 95% confidence intervals were calculated using Poisson regression to determine risk factors for carriage. Results A total of 1837 oropharyngeal specimens were obtained from 1478 unique participants. Overall carriage prevalence was 12.7–14.6% during the two survey rounds, with 1.8–2.6% for capsular genotype B, 0.9–1.0% for capsular genotypes C, W, or Y, and 9.9–10.8% for nongroupable strains by rt-PCR. Meningococcal carriage was associated with being male, smoking, party or club attendance, recent antibiotic use (inverse correlation), and recent respiratory infections. Conclusions In this university setting, the majority of meningococcal carriage was due to nongroupable strains, followed by serogroup B. Further evaluation is needed to understand the dynamics of serogroup B carriage and disease among university students.

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“…For eight cases, N. meningitidis serogroup was determined by slide agglutination at the state public health laboratory and confirmed by real-time polymerase chain reaction (rt-PCR) at CDC [2–4]. Serogroup determination and PorA typing for one case was performed in Greece using PCR.…”

Section: Methodsmentioning

confidence: 99%

First Use of a Serogroup B Meningococcal Vaccine in the US in Response to a University Outbreak

et al. 2015

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Background In 2013–2014, an outbreak of serogroup B meningococcal disease occurred among persons linked to a New Jersey university (University A). In the absence of a licensed serogroup B meningococcal (MenB) vaccine in the US, the Food and Drug Administration authorized use of an investigational MenB vaccine to control the outbreak. An investigation of the outbreak and response was undertaken to determine the population at risk and assess vaccination coverage. Methods The epidemiologic investigation relied on compilation and review of case and population data, laboratory typing of meningococcal isolates, and unstructured interviews with university staff. Vaccination coverage data were collected during the vaccination campaign held under an expanded access Investigational New Drug protocol. Results Between March 25, 2013 and March 10, 2014, 9 cases of serogroup B meningococcal disease occurred in persons linked to University A. Laboratory typing results were identical for all 8 isolates available. Through May 14, 2014, 89.1% coverage with the two-dose vaccination series was achieved in the target population. From the initiation of MenB vaccination through February 1, 2015, no additional cases of serogroup B meningococcal disease occurred in University A students. However, the 9th case occurred in March 2014 in an unvaccinated close contact of University A students. Conclusions No serogroup B meningococcal disease cases occurred in persons who received 1 or more doses of 4CMenB vaccine, suggesting 4CMenB may have protected vaccinated individuals from disease. However, the 9th case demonstrates that carriage of serogroup B Nesisseria meningitidis among vaccinated persons was not eliminated.

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sodC-Based Real-Time PCR for Detection of Neisseria meningitidis

Cited by 81 publications

References 40 publications

Comparison of Phenotypic and Genotypic Approaches to Capsule Typing of Neisseria meningitidis by Use of Invasive and Carriage Isolate Collections

Comparison of Phenotypic and Genotypic Approaches to Capsule Typing of Neisseria meningitidis by Use of Invasive and Carriage Isolate Collections

Meningococcal carriage among a university student population – United States, 2015

First Use of a Serogroup B Meningococcal Vaccine in the US in Response to a University Outbreak

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