Work

Application and evaluation of a rapid detection method based on two-dimensional PCR technology for hypervirulent Klebsiella pneumoniae

Year: 2025

Type: article

Abstract: Objectives Hypervirulent K. pneumoniae (hvKp) is an emerging pathogen that is more virulent than classical K. pneumoniae (cKp). This study aimed to develop an economical, high-throughput, and accurate two-dimensional polymerase chain reaction (2D-PCR) assay for the rapid detection of hvKp. Materials and methods Recombinant plasmids containing the iucA, peg-344, rmpA2, and rmpA virulence genes were constructed and used for assessing the sensitivity and specificity of the 2D-PCR. Clinical samples (n = 105) were collected and evaluated the performance of the 2D-PCR to comparison with conventional PCR methods. Results The minimum detection limit of the 2D-PCR assay for iucA, peg-344, rmpA2, and rmpA were 103, 102, 103, and 103 copies/μL, respectively. Additionally, the concordance rates between the 2D-PCR and conventional PCR for detecting iucA, peg-344, rmpA2, and rmpA were all over 95%. The analysis revealed a sensitivity of 100.0% and a specificity of 96.2% when compared to conventional PCR. Conclusion A 2D-PCR-based multiplex method for virulence genes of hvKp was successfully developed, demonstrating its outstanding features of high specificity, high sensitivity, and high throughput capability. This method could be used for the rapid diagnosis of infectious diseases caused by hvKp in clinical settings.

Source: Frontiers in Microbiology

DOI: 10.3389/fmicb.2025.1554660

Authors: Wenwen Zhu, Yiting Wang, Xin Jiang, Yan Zhao

Institutions: Shanghai Medical College of Fudan University, Fudan University, Jinshan Hospital of Fudan University

Language: English

Topic: Bacterial Identification and Susceptibility Testing

Subfield: Clinical Biochemistry

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $3,801