Rapid identification of Streptococcus pneumoniae serotypes by cpsB gene-based sequetyping combined with multiplex PCR

Zhou, Mu; Wang, Zi-Ran; Li, Yanbing; Kudinha, Timothy; Wang, Jian; Wang, Yao; Xiao, Meng; Xu, Yingchun; Liu, Zhengyin; Hsueh, Po‐Ren

doi:10.1016/j.jmii.2021.11.004

Cited by 6 publications

(7 citation statements)

References 31 publications

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“…Test results of the simulated clinical samples suggested that the assay most probably could meet clinical laboratory requirement. Overall, these performance characteristics are similar to those observed in assays developed for rapid identi cation of SARS-CoV-2 variants ( 22), E.coli serotyping (21) and Streptococcuspneumoniae serotyping (23). In the multicenter clinical study, EFMAtyping assay had shown perfect concordance with the WGS serological method.…”

Section: Discussionsupporting

confidence: 69%

“…Multicolor melting curve analysis (MMCA) is a homogeneous multiplex analytical system with uorophore color and melting temperature (Tm) value as the two-dimensional label (11). MMCA has been successfully used for simultaneous identi cation of 30 common Salmonella serotypes or 11 clinically commonest V. parahaemolyticus serotypes by a multiplex ligation reaction based on probe MCA (12,13), for identi cation of clinically relevant Mycobacterium species (14), and for identi cation of 92 pneumococcal serotypes (15). In this study, we developed a revised version of MMCA, EFMAtyping (enteric fever MeltArray typing), a single PCR reaction assay which is more speci c for identifying both typhoidal and paratyphoidal Salmonella, and its performance was further evaluated by testing 551 clinical Salmonella isolates collected over a 17-year period (2001-2017) in Shenzhen city.…”

Section: Introductionmentioning

confidence: 99%

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Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Jiang,

Cai

et al. 2024

Preprint

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Rapid and accurate identification of Salmonella enterica serotypes Typhi and Paratyphi (A, B and C), the causal agents of enteric fever, is critical for timely treatment, case management and evaluation of health policies in low and middle-income countries where the disease still remains a serious public health problem. The present study describes the development of a multiplex assay (EFMAtyping) for simultaneous identification of pathogens causing typhoid and paratyphoid fever in a single reaction by the MeltArray approach, which could be finished within 2.5 h. Seven specific genes were chosen for differentiation of typhoidal and nontyphoidal Salmonella. All gene targets were able to be detected by the EFMAtyping assay, with expected Tm values and without cross-reactivity to other relevant Salmonella serovars. The limit of detection (LOD) for all gene targets was 50 copies per reaction. The LOD reached 102-103 CFU/ml for each pathogen in simulated clinical samples. The largest standard deviation value for mean Tm was below 0.5°C. This newly developed EFMAtyping assay was further evaluated by testing 551 clinical Salmonella isolates, corroborated in parallel by the traditional Salmonella identification workflow, and serotype prediction was enabled by whole-genome sequencing. Compared to the traditional method, our results exhibited 100% of specificity and greater than 96% of sensitivity with a kappa correlation ranging from 0.96 to 1.00. Thus, the EFMAtyping assay provides a rapid, high throughput, and promising tool for public health laboratories to monitor typhoid and paratyphoid fever.

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Section: Discussionsupporting

confidence: 69%

Section: Introductionmentioning

confidence: 99%

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Jiang,

Cai

et al. 2024

Preprint

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“…If the specimen’s initial three reactions were negative, it was subjected to eight sequential multiplex reactions as specified by CDC ( Jin et al 2016 ). In another study, a multiplex system with eight reactions successfully identified 73.3% of S. pneumoniae serotypes in the study collection; nevertheless, identifying the serotypes 17A, 24F, 28A, 28F, 29, and 33B was unsuccessful primarily because of the limited target primers in the multiplex PCR ( Zhou et al 2021 ).…”

Section: Serotypingmentioning

confidence: 99%

“…It was found that cpsB could not be amplified for serotypes 25F, 37, 38, 39, and 43 ( Mauffrey et al 2017 ). There is still significant homology concerning cpsB from some distinct serotypes, which can only be identified as a combined form of serotypes or serogroup, e.g., 6C-6D or 13-20A-20B ( Zhou et al 2021 ).…”

Section: Serotypingmentioning

confidence: 99%

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The Molecular Approaches and Challenges of Streptococcus pneumoniae Serotyping for Epidemiological Surveillance in the Vaccine Era

Abdul Rahman,

Mohd Desa,

Masri

et al. 2023

Polish Journal of Microbiology

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Streptococcus pneumoniae (pneumococcus) belongs to the Gram-positive cocci. This bacterium typically colonizes the nasopharyngeal region of healthy individuals. It has a distinct polysaccharide capsule – a virulence factor allowing the bacteria to elude the immune defense mechanisms. Consequently, it might trigger aggressive conditions like septicemia and meningitis in immunocompromised or older individuals. Moreover, children below five years of age are at risk of morbidity and mortality. Studies have found 101 S. pneumoniae capsular serotypes, of which several correlate with clinical and carriage isolates with distinct disease aggressiveness. Introducing pneumococcal conjugate vaccines (PCV) targets the most common disease-associated serotypes. Nevertheless, vaccine selection pressure leads to replacing the formerly dominant vaccine serotypes (VTs) by non-vaccine types (NVTs). Therefore, serotyping must be conducted for epidemiological surveillance and vaccine assessment. Serotyping can be performed using numerous techniques, either by the conventional antisera-based (Quellung and latex agglutination) or molecular-based approaches (sequetyping, multiplex PCR, real-time PCR, and PCR-RFLP). A cost-effective and practical approach must be used to enhance serotyping accuracy to monitor the prevalence of VTs and NVTs. Therefore, dependable pneumococcal serotyping techniques are essential to precisely monitor virulent lineages, NVT emergence, and genetic associations of isolates. This review discusses the principles, associated benefits, and drawbacks of the respective available conventional and molecular approaches, and potentially the whole genome sequencing (WGS) to be directed for future exploration.

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Identification of pneumococcal serotypes with individual recognition of vaccine types by a highly multiplexed real-time PCR-based MeltArray approach

Zhou,

Che,

Wang

et al. 2024

Journal of Microbiology, Immunology and Infection

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Rapid identification of Streptococcus pneumoniae serotypes by cpsB gene-based sequetyping combined with multiplex PCR

Cited by 6 publications

References 31 publications

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

Rapid and Specific Differentiation of Salmonella enterica serotypes Typhi and Paratyphi by Multicolor Melting Curve Analysis

The Molecular Approaches and Challenges of Streptococcus pneumoniae Serotyping for Epidemiological Surveillance in the Vaccine Era

Identification of pneumococcal serotypes with individual recognition of vaccine types by a highly multiplexed real-time PCR-based MeltArray approach

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