Journal Article

Use of a rapid recombinase-aided amplification assay for Mycoplasma pneumoniae detection

Li, Shaoli, Jiang, Tingting, Feng, Yanling, Cui, Jinghua, Xue, Guanhua, Zhao, Hanqing, Yuan, Jing, Yan, Chao

2020

Overview

Background Mycoplasma pneumoniae is one of the most common causative pathogens of community-acquired pneumonia (CAP), accounting for as many as 30–50% of CAP during peak years. An early and rapid diagnostic method is key for guiding clinicians in their choice of antibiotics . Methods The recombinase-aided amplification (RAA) assay is a recently developed, rapid detection method that has been used for the detection of several pathogens. The assays were performed in a one-step single tube reaction at 39° Celsius within 15–30 min. In this study, we established an RAA assay for M. pneumoniae using clinical specimens for validation and commercial real-time PCR as the reference method. Results The analytical sensitivity of the RAA assay was 2.23 copies per reaction, and no cross-reactions with any of the other 15 related respiratory bacterial pathogens were observed. Compared with the commercial real-time PCR assay used when testing 311 respiratory specimens, the RAA assay obtained 100% sensitivity and 100% specificity with a kappa value of 1. Conclusions These results demonstrate that the proposed RAA assay will be of benefit as a faster, sensitive, and specific alternative tool for the detection of M. pneumoniae .

• Publisher: BioMed Central,BioMed Central Ltd,Springer Nature B.V,BMC
• Subject: Adolescent / Amplification / Antibiotics / Assaying / Bacterial infections / Bronchitis / Child / Child, Preschool / Community-Acquired Infections - microbiology / Deoxyribonucleic acid / Detection / Diagnosis / Diagnostic bacteriology / Diagnostic systems / DNA / DNA polymerase / Enzymes / Epidemics / Female / Genetic testing / Health aspects / Humans / Infant / Infant, Newborn / Infectious Diseases / Internal Medicine / Male / Medical Microbiology / Medicine / Medicine & Public Health / Methods / Molecular diagnostic technique / Mycoplasma pneumoniae / Mycoplasma pneumoniae - genetics / Nucleic Acid Amplification Techniques - methods / Parasitology / Pathogens / Pediatric research / Pediatrics / Performance evaluation / Plasmids / Pneumonia / Pneumonia, Mycoplasma - microbiology / Real time / Recombinase / Recombinase-aided amplification / Recombinases / Recombinases - genetics / Reproducibility of Results / Research Article / Risk factors / Sensitivity analysis / Sensitivity and Specificity / Statistical analysis / Streptococcus infections / Tropical Medicine / Tuberculosis and other mycobacterial diseases