Establishment of a rapid detection method for Mycoplasma pneumoniae based on RPA-CRISPR-Cas12a technology.

Clin Chim Acta

The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui, China, 230022; Anhui Public Health Clinical Center, Hefei, Anhui, China, 230012. Electronic address:

Published: January 2025

Mycoplasma pneumoniae can cause respiratory infections and pneumonia, posing a serious threat to the health of children and adolescents. Early diagnosis of Mycoplasma pneumoniae infection is crucial for clinical treatment. Currently, diagnostic methods for Mycoplasma pneumoniae infection include pathogen detection, molecular biology techniques, and bacterial culture, all of which have certain limitations. Here, we developed a rapid, simple, and accurate detection method for Mycoplasma pneumoniae that does not rely on large equipment or complex operations. This technology combines the CRISPR-Cas12a system with recombinase polymerase amplification (RPA), allowing the detection results to be observed through fluorescence curves and immunochromatographic lateral flow strips.It has been validated that RPA-CRISPR/Cas12a fluorescence analysis and RPA-CRISPR/Cas12-immunochromatographic exhibit no cross-reactivity with other common pathogens, and The established detection limit was ascertained to be as low as 10 copies/µL.Additionally, 49 clinical samples were tested and compared with fluorescence quantitative polymerase chain reaction, demonstrating a sensitivity and specificity of 100%. This platform exhibits promising clinical performance and holds significant potential for clinical application, particularly in settings with limited resources, such as clinical care points or resource-constrained areas.

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.cca.2024.119906DOI Listing

Publication Analysis

Top Keywords

mycoplasma pneumoniae
20
detection method
8
method mycoplasma
8
pneumoniae infection
8
detection
5
mycoplasma
5
pneumoniae
5
clinical
5
establishment rapid
4
rapid detection
4

Similar Publications

[Analysis of drug-resistance mutations and treatment of mycoplasma pneumonia in children in Hunan Province in 2023].

Zhonghua Yu Fang Yi Xue Za Zhi

December 2024

Department of Laboratory Medicine, The Second Xiangya Hospital, Central South University, Changsha410011, China.

To investigate the drug-resistance mutations and treatment of hospitalized children with Mycoplasma pneumoniae pneumonia (MPP) in Hunan Province. Children with pneumonia, who were hospitalized in the pediatric ward of the Second Xiangya Hospital of Central South University from January 1, 2023, to December 31, 2023, were enrolled in this study, and their clinical data was also collected. The targeted next-generation sequencing (tNGS) was used to detect Mycoplasma pneumoniae (MP) infection and drug-resistance mutations, and the drug-resistance and treatment in children with MPP were also analyzed.

View Article and Find Full Text PDF

CRISPR-Cas12a2-based rapid and sensitive detection system for target nucleic acid.

Int J Biol Macromol

December 2024

Shanghai Institute of Infectious Disease and Biosecurity, Fudan University, Shanghai 200032, China; Department of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Fudan University, Shanghai 20032, China. Electronic address:

Infectious diseases are extremely important public health issues, where the design of effective, rapid, and convenient detection platforms is critical. In this study, we coupled SuCas12a2, a novel Cas12 family RNA-targeting nuclease, with conventional PCR and recombinase polymerase amplification (RPA), respectively, to develop novel detection approaches, named PCR-SuCas12a2 and RPA-SuCas12a2. SuCas12a2 possesses collateral cleavage activity and cuts the additional single-stranded RNA (ssRNA) added to the reaction system once the ternary complex RNA-SuCas12a2-CRISPR RNA (crRNA) is formed.

View Article and Find Full Text PDF

The contribution of the lung microbiota to pneumonia in children of varying severity remains poorly understood. This study utilized metagenomic next-generation sequencing (mNGS) technology to elucidate the characteristics of lung microbiota and their association with disease severity. This retrospective study analyzed bronchoalveolar lavage fluid (BALF) mNGS data of 92 children diagnosed with pneumonia between January 2021 and July 2022.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!