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

Zhou, Shujuan; Che, Jie; Wang, Xuran; Lin, Yong; Niu, Jianjun; Liang, Weitong; Xu, Li; Zhang, Maojun; Liao, Yiqun; Shao, Zhujun; Li, Qingge

doi:10.1016/j.jmii.2023.10.008

Cited by 2 publications

(1 citation statement)

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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%

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%