Serotyping Pneumococcal Meningitis Cases in the African Meningitis Belt by Use of Multiplex PCR with Cerebrospinal Fluid

Lafourcade, Berthe-Marie Njanpop; Sanou, Oumarou; Linden, Mark van der; Levina, Natalia; Karanfil, Meryem; Yaro, Seydou; Tamekloe, Tsidi Agbeko; Mueller, Judith E.

doi:10.1128/jcm.01402-09

Cited by 23 publications

(32 citation statements)

References 11 publications

(9 reference statements)

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“…In addition, 169 isolates from Brazil, India, Kenya, Mongolia, Mozambique, Nepal, Peru, and Thailand were included. Forty-three isolates of 15 related species, which included Streptococcus pseudopneumoniae (10), Streptococcus gordonii (6) [18]) were tested. DNA was extracted using the Qiagen DNA minikit (Qiagen Inc., Valencia, CA) (18).…”

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confidence: 99%

Sequential Triplex Real-Time PCR Assay for Detecting 21 Pneumococcal Capsular Serotypes That Account for a High Global Disease Burden

et al. 2013

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cWe developed and validated a real-time PCR assay consisting of 7 triplexed reactions to identify 11 individual serotypes plus 10 small serogroups representing the majority of disease-causing isolates of Streptococcus pneumoniae. This assay targets the 13 serotypes included within the 13-valent conjugate vaccine and 8 additional key serotypes or serogroups. Advantages over other serotyping assays are described. The assay will be expanded to 40 serotypes/serogroups. We will provide periodic updates at our protocol website.

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confidence: 99%

Sequential Triplex Real-Time PCR Assay for Detecting 21 Pneumococcal Capsular Serotypes That Account for a High Global Disease Burden

et al. 2013

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“…While other studies have used oligonucleotide permutations to account for geographical differences in S. pneumoniae serotype distribution (USA, Latin America, Africa, and Asia), [17,24,40] this study showed that oligonucleotides permutations did not affect the performance characteristics of PCR-based serotyping. This study does not preclude additional validation if different permutations are used, or use of ongoing quality assurance controls.…”

Section: Discussionmentioning

confidence: 69%

“…[15] More recently, molecular methods for serotype deduction like conventional multiplex PCRs (cmPCR) have been developed, and are widely used from DNA extracted from S. pneumoniae isolates or clinical specimens. [16][17][18][19][20][21][22][23][24][25][26][27][28] Since cmPCR have a limited number of serotypes in each PCR reaction, they have been designed to target the most prevalent serotypes causing IPD, often in sequential reactions. [21][22][23][24][25][26] However, sequential PCR may not be the most cost effective strategy to identify S. pneumo-niae serotypes that are vaccine-preventable.…”

Section: Introductionmentioning

confidence: 99%

Refining PCR-based serotyping for detection of vaccine-preventable Streptococcus pneumoniae

Lang

Gillis

ElSherif

et al. 2016

JER

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Conventional multiplex PCR (cmPCR) reactions have been developed to monitor the most predominant serotypes of Streptococcus pneumoniae causing invasive pneumococcal disease (IPD). Since cmPCR assigns serotypes based on differences in the capsule biosynthesis (cps) loci, DNA extracted from clinical specimens can be used directly to monitor changes in serotype distribution and assess the impact of pneumococcal vaccines. Given that cmPCR can require up to eight reactions to assign a serotype, testing is often conducted in sequential algorithms. Sequential cmPCR reactions; however, may not be the most cost effective strategy to determine whether a S. pneumoniae serotype is vaccine-preventable. This study used oligonucleotide permutations in a modified set of cmPCR reactions (termed cmPCRmod) to reduce the number of PCR reactions required to identify S. pneumoniae serotypes covered by the 7-and 13-valent pneumococcal conjugate vaccines (PCV7 and PCV13, respectively) and the 23-valent pneumococcal polysaccharide vaccine (PPV23). While oligonucleotide permutations have previously been reported for regional differences in serotype distribution, the impact on assay performance had not been assessed. This study demonstrated that equivalent analytical sensitivity and specificity was seen when comparing cmPCR and cmPCRmod, and 100% concordance was seen when 308 clinical isolates of S. pneumoniae were evaluated. Compared to cmPCR, cmPCRmod reduced the number and reactions required to detect serotypes covered by PCV7, PCV13, and PPV23. This study demonstrated that conventional multiplex reactions can be reformulated for more efficient detection of vaccine-preventable serotypes, without compromising test performance characteristics. As such, cmPCRmod reactions could provide significant cost savings for large surveillance studies.

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“…In contrast, Quellung reaction is subjective and a high level of experience is required to assign the correct serotype. Different authors have adapted the original multiplex PCR assay developed by Pai et al (29) and recommended by the CDC (1,3,11,12,18,26,28,31). All authors agree that multiplex PCR is an accurate, simple, and economical method which can be used for determining capsular type.…”

Section: Discussionmentioning

confidence: 99%

Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis

et al. 2012

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The purpose of this study was to develop a high-throughput method for the identification of pneumococcal capsular types. Multiplex PCR combined with fragment analysis and automated fluorescent capillary electrophoresis (FAF-mPCR) was utilized. Only 11 results were discordant with the Quellung reaction. However, latex reaction and Quellung results of the second reference laboratory agreed with FAF-mPCR for 9 of these 11 strains (82%). Therefore, we considered that only 2 of 394 strains (0.5%) were not properly characterized by the new assay. The automation of the process allowed the typing of 30 isolates in a few hours with a lower cost than that of the Quellung reaction. These results indicate that FAF-mPCR is a good method to determine the capsular serotype of Streptococcus pneumoniae.

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Serotyping Pneumococcal Meningitis Cases in the African Meningitis Belt by Use of Multiplex PCR with Cerebrospinal Fluid

Cited by 23 publications

References 11 publications

Sequential Triplex Real-Time PCR Assay for Detecting 21 Pneumococcal Capsular Serotypes That Account for a High Global Disease Burden

Sequential Triplex Real-Time PCR Assay for Detecting 21 Pneumococcal Capsular Serotypes That Account for a High Global Disease Burden

Refining PCR-based serotyping for detection of vaccine-preventable Streptococcus pneumoniae

Rapid and Easy Identification of Capsular Serotypes of Streptococcus pneumoniae by Use of Fragment Analysis by Automated Fluorescence-Based Capillary Electrophoresis

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