Evaluation of Pneumococcal Serotyping by Multiplex PCR and Quellung Reactions

Richter, Sandra; Heilmann, Kristopher P.; Dohrn, Cassie L.; Riahi, Fathollah; Diekema, Daniel J.; Doern, Gary V.

doi:10.1128/jcm.01876-13

Cited by 24 publications

(15 citation statements)

References 13 publications

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“…Since, similar to our results, the serotype identified by the UAD assay had been shown in a previous study to correlate 100% with the serotypes from blood cultures in patients with bacteremic CAP (27), alternative explanations need to be considered for the discrepancy from carriage serotypes, not taking into account the possibility of technical error or mislabeling (41). This finding might reflect a previous infection rather than an incorrect detection of the carriage serotype.…”

Section: Discussionsupporting

confidence: 75%

Multiplex Urinary Antigen Detection for 13 Streptococcus pneumoniae Serotypes Improves Diagnosis of Pneumococcal Pneumonia in South African HIV-Infected Adults

et al. 2017

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A serotype-specific urinary antigen detection (UAD) assay for 13 serotypes included in the pneumococcal conjugate vaccine (PCV13) was recently reported as a useful diagnostic tool for pneumococcal pneumonia. We aimed to assess the diagnostic accuracy of the UAD in HIV-infected South African adults. Urine specimens from a well-defined cohort of HIV-infected South African adults with pneumonia were evaluated retrospectively in the UAD assay. Pneumonia was considered pneumococcal if either sputum Gram stain, sputum culture, blood culture, or the immunochromatographic (ICT) BinaxNow S. pneumoniae test (composite diagnostic) was positive. Among 235 enrolled pneumonia patients, the UAD assay was more frequently positive (104 [44.3%]) than the composite diagnostic (71 [30.2%]; P Ͻ 0.001) and increased the pneumococcal etiology from 30.2% by an additional 22.6% to 52.8%. The UAD assay detected more pneumococcal etiologies (45.0%) than the serotype-independent ICT (23.4%, P Ͻ 0.001). UAD identified 6/7 patients with PCV13 serotype bacteremia without misclassification of bacteremia episodes due to non-PCV13 serotypes. UAD was positive for 5.1% of asymptomatic HIV-infected persons, with higher rates among those with nasopharyngeal carriage. Concordance between serotypes identified by UAD and by Quellung reaction and PCR serotyping was 70/86 (81.4%). UAD identified the dominant serotype in multiple serotype carriage. This study confirms the utility of the UAD assay for HIV-infected adults comparing favorably with other diagnostic tests. A highly valent UAD may become a new standard for detection of pneumococcal pneumonia in adults. Prior to PCV introduction, at least 53% of pneumonia cases were due to pneumococci in HIVinfected South African adults.

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Section: Discussionsupporting

confidence: 75%

Multiplex Urinary Antigen Detection for 13 Streptococcus pneumoniae Serotypes Improves Diagnosis of Pneumococcal Pneumonia in South African HIV-Infected Adults

et al. 2017

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“…In addition, any isolate that could not be serotyped was tested using an AccuProbe S. pneumoniae culture identification test (GenProbe). Capsular serotypes were determined by using a multiplex PCR targeting the most common serotypes (8,9) and the Quellung reaction with type-specific antisera (Statens Serum Institut) where necessary.…”

Section: Methodsmentioning

confidence: 99%

Changes in Pneumococcal Serotypes and Antimicrobial Resistance after Introduction of the 13-Valent Conjugate Vaccine in the United States

Richter

Diekema

Heilmann

et al. 2014

Antimicrob Agents Chemother

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143

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“…To assess S. pneumoniae serotype distribution, serotyping methods using Quellung reactions or latex agglutination are often used, but these require viable pneumococci and large panels of antisera (Austrian, 1976;Lovgren et al, 1998;Siira et al, 2012;Ortika et al, 2013). Molecular methods to deduce S. pneumoniae serotypes, such as conventional multiplex PCRs (cmPCR) and realtime multiplex PCRs (rmPCR), have been shown to be a more practical alternative for large epidemiological studies, and have been applied to bacterial isolates and clinical specimens (Pai et al, 2006;Dias et al, 2007, Morais et al, 2007Azzari et al, 2008;Lafourcade et al, 2010;Jourdain et al, 2011;Pimenta et al, 2013;Richter et al, 2013;Shakrin et al, 2013). This study evaluated the feasibility of PCR-based detection and serotyping of pneumococci directly in nasopharyngeal (NP) swabs routinely collected for viral studies to add to the repertoire of non-culture methods used for pneumococcal surveillance.…”

Section: Introductionmentioning

confidence: 99%

Detection and prediction of Streptococcus pneumoniae serotypes directly from nasopharyngeal swabs using PCR

Lang

McNeil

Hatchette

et al. 2015

Journal of Medical Microbiology

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Monitoring Streptococcus pneumoniae serotype distribution is important to assess the impact and effectiveness of pneumococcal vaccine programs. With the challenges of Quellung serotyping, PCR-based serotype prediction is increasingly being used for large-scale epidemiological studies. This study used real-time (RT)-PCR targeting the genes encoding autolysin (lytA) and capsular biosynthesis gene A (cpsA) of S. pneumoniae in nucleic acids extracted directly from nasopharyngeal (NP) swabs submitted for viral studies. If the specimen was lytA or cpsA PCR-positive, then serotype prediction was performed on the same nucleic acid using eight conventional multiplex PCRs (cmPCRs) and seven real-time multiplex PCRs (rmPCRs). Of 1770 NP swabs, 132 (7.5 %) were lytA-positive and 122 (6.9 %) were positive for both targets (lytA and cpsA). Of the 122 lytA + cpsA + specimens, a serotype could be assigned in 52 (41.8 %) using cmPCR alone and the yield was increased to 70 (57.4 %) with the addition of rmPCR. Based on sensitivity, incremental yield and more efficient workflow, an algorithm was proposed where lytA and cpsA RT-PCR screening was followed by serotype deduction using rmPCR and a modified set of four instead of eight cmPCRs. This algorithm was validated for use on NP swabs, and the distribution of S. pneumoniae serotypes deduced from this approach showed good concordance with those obtained with cultured isolates serotyped by Quellung and PCR. Overall, molecular detection and serotyping of S. pneumoniae from NP swabs was found to be a valuable tool to assess S. pneumoniae colonization and monitor trends in serotype distribution.

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Evaluation of Pneumococcal Serotyping by Multiplex PCR and Quellung Reactions

Cited by 24 publications

References 13 publications

Multiplex Urinary Antigen Detection for 13 Streptococcus pneumoniae Serotypes Improves Diagnosis of Pneumococcal Pneumonia in South African HIV-Infected Adults

Multiplex Urinary Antigen Detection for 13 Streptococcus pneumoniae Serotypes Improves Diagnosis of Pneumococcal Pneumonia in South African HIV-Infected Adults

Changes in Pneumococcal Serotypes and Antimicrobial Resistance after Introduction of the 13-Valent Conjugate Vaccine in the United States

Detection and prediction of Streptococcus pneumoniae serotypes directly from nasopharyngeal swabs using PCR

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