Evaluation of Commercial Molecular Diagnostic Methods for Detection and Determination of Macrolide Resistance in Mycoplasma pneumoniae

Abstract

We evaluated six commercial molecular tests targeting Mycoplasma pneumoniae, namely, the BioFire FilmArray respiratory panel (RP), the Meridian Alethia Mycoplasma Direct, the GenMark ePlex respiratory pathogen panel (RPP), the Luminex NxTAG RPP, the ELITech ELITe InGenius Mycoplasma MGB research use only (RUO) PCR, and the SpeeDx Resistance Plus MP assays. Laboratory-developed PCR assays at the University of Alabama at Birmingham and the Centers for Disease Control and Prevention were used as reference standards. Among 428 specimens, 212 were designated confirmed positives for M. pneumoniae. The highest clinical sensitivities were found with the InGenius PCR (99.5%) and the FilmArray RP (98.1%). The Resistance Plus MP identified 93.3% of the confirmed-positive specimens, whereas 83.6, 64.6, and 55.7% were identified by the ePlex RPP, NxTAG RPP, and Mycoplasma Direct assays, respectively. There was no significant difference between the sensitivity of the reference methods and that of the FilmArray RP and InGenius assays, but the remaining four assays detected significantly fewer positive specimens (P < 0.05). Specificities of all assays were 99.5 to 100%. The Resistance Plus MP assay detected macrolide resistance in 27/33 specimens, resulting in a sensitivity of 81.8%. This study provides the first large-scale comparison of commercial molecular assays for detection of M. pneumoniae in the United States and identified clear differences among their performance. Additional studies are necessary to explore the impact of various test performances on patient outcome.

Point-of-care molecular diagnosis of Mycoplasma pneumoniae including macrolide sensitivity using quenching probe polymerase chain reaction

“…Commercial molecular assays for detection of M . pneumoniae are listed in Table 3 [ 19 , 39 – 49 ]. The major features of the Smart Gene ® system and other assays are summarized as follows.…”

“…Nucleic acid amplification techniques (NAATs) including quantitative polymerase chain reaction (qPCR) assay, multiplex qPCR assay and loop-mediated isothermal amplification (LAMP) have been increasingly used for identification of respiratory pathogens including M . pneumoniae in clinical specimens due to their high levels of sensitivity and specificity [ 18 , 19 ]. QPCR and multiplex qPCR assays usually require high-performance instruments and well-equipped laboratories, and these assays can detect macrolide-resistant point mutations at domain V of the 23S rRNA gene of M .…”

Mycoplasma pneumoniae Pleural Effusion in Adults

“…In regions where the development of macrolide resistance is suspected based on local epidemiology research, such as most of Asia, alternative therapy (doxycycline or fluoroquinolone) should be considered in patients with severe or refractory diseases [ 71 ]. Macrolide resistance tests have been developed using a molecular method and are expected to be conducted more frequently [ 72 ].…”

Development of a Rapid and Efficient RPA-CRISPR/Cas12a Assay for Mycoplasma pneumoniae Detection

“…Mycoplasma pneumoniae (MP) is a major causative agent of community-acquired pneumonia in humans, especially children and adolescents, accounting for up to 40% of cases ( Leal et al, 2020 ; Xue et al, 2020 ; Kakiuchi et al, 2021 ). MP infections occur throughout the year with epidemic peaks at intervals of 3-7 years ( Atkinson et al, 2008 ; Rivaya et al, 2020 ; Jiang et al, 2021 ).…”

“…The high morbidity of MP infections reinforces the need for timely diagnosis. While culturing of MP is time-consuming and relatively insensitive due to its slow growth and fastidious nature ( Leal et al, 2020 ). Reliable results of serological test rely on a single high antibody titer, or ≥ 4-fold increase in antibody titers of paired acute-phase and convalescent-phase serums, nevertheless, the former often exhibits false-negative result in the early stages of MP infection due to multiple factors, and the later requires 2-4 weeks of monitoring, which delays diagnosis and complicates the clinical course ( She et al, 2010 ; Kishaba, 2016 ).…”