Detection of Meningococcal Carriage by Culture and PCR of Throat Swabs and Mouth Gargles

Jordens, J. Zoe; Williams, Jeannette N.; Jones, Graeme R.; Heckels, John E.

doi:10.1128/jcm.40.1.75-79.2002

Cited by 39 publications

(40 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…All N. meningitidis isolates were tested using an rt-PCR assay targeting the porA (encoding the class 1 or PorA protein) and ctrA genes (76,77). All isolates (regardless of porA and ctrA results) were evaluated further in rt-PCR assays specific for the eight capsule serogroups, as described by Rojas et al (68).…”

Section: Meningococcalmentioning

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

View full text Add to dashboard Cite

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 ...

show abstract

Section: Meningococcalmentioning

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

View full text Add to dashboard Cite

show abstract

“…verse primer ( Table 1) described by Corless et al (4) and demonstrated to amplify a region of the ctrA gene from all meningococcal serogroups (4,11).…”

mentioning

confidence: 99%

PCR-Based Assay for Detection of Neisseria meningitidis Capsular Serogroups 29E, X, and Z

2004

View full text Add to dashboard Cite

show abstract

“…Genes specific for N. meningitidis detection and genogrouping. ctrA or porA genes have been used individually to identify N. meningitidis (15,16). However, we found that our primers based on the ctrA sequence produced PCR products from both N. meningitidis and N. lactamica, and those based on the porA sequence gave PCR products from both N. meningitidis and N. gonorrhoeae (data not shown).…”

Section: Methodsmentioning

confidence: 56%

“…Infections with N. meningitidis are a significant cause of mortality and morbidity in young children and adolescents (16,23). The bacterial capsular polysaccharide (CPS) is the most important virulence factor for N. meningitidis (14,20,26,28,32).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Development of a Multiplex PCR Assay for Detection and Genogrouping of Neisseria meningitidis

et al. 2012

View full text Add to dashboard Cite

Neisseria meningitidis is a leading pathogen of epidemic bacterial meningitis and fulminant sepsis worldwide. Twelve different N. meningitidis serogroups have been identified to date based on antigenic differences in the capsular polysaccharide. However, more than 90% of human cases of N. meningitidis meningitis are the result of infection with just five serogroups, A, B, C, W135, and Y. Efficient methods of detection and genogrouping of N. meningitidis isolates are needed, therefore, in order to monitor prevalent serogroups as a means of disease control and prevention. The capsular gene complex regions have been sequenced from only seven out of the 12 serogroups. In this study, the capsular gene complexes of the remaining five serogroups were sequenced and analyzed. Primers were designed that were specific for N. meningitidis species and for the 12 individual serogroups, and a multiplex PCR assay using these specific primers was developed for N. meningitidis detection and genogrouping. The assay was tested using 15 reference strains covering all 12 serogroups, 143 clinical isolates, and 21 strains from closely related species or from species that cause meningitis. The assay could detect N. meningitidis serogroups and was shown to be specific, with a detection sensitivity of 1 ng of genomic DNA (equivalent to ϳ4 ؋ 10 5 genomes) or 3 ؋ 10 5 CFU/ml in noncultured mock cerebrospinal fluid (CSF) specimens. This study, therefore, describes for the first time the development of a molecular protocol for the detection of all N. meningitidis serogroups. This multiplex PCR-based assay may have use for the clinical diagnosis and epidemiological surveillance of N. meningitidis.

show abstract

Detection of Meningococcal Carriage by Culture and PCR of Throat Swabs and Mouth Gargles

Cited by 39 publications

References 12 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

PCR-Based Assay for Detection of Neisseria meningitidis Capsular Serogroups 29E, X, and Z

Development of a Multiplex PCR Assay for Detection and Genogrouping of Neisseria meningitidis

Contact Info

Product

Resources

About