Rapid Detection of the Macrolide Sensitivity of Pneumonia-Causing Mycoplasma pneumoniae Using Quenching Probe Polymerase Chain Reaction (GENECUBE®)

Ito, Yutaka; Iwashima, Satoru; Hayano, S.; Nishio, Tomohiro; Shiozawa, R; Yata, Soichiro; Kubota, Toshiko; Kubota, Akira; Uemura, Keiichi

doi:10.1007/s40291-018-0360-x

Cited by 13 publications

(12 citation statements)

References 36 publications

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“…GENECUBE Mycoplasma-assisted clinical diagnosis can contribute to the optimization of the selection of antibiotics and the improvement of the clinical course of a disease [17]. Supporting these findings, the results obtained by using the automated molecular system and 23S rRNA sequencing were completely identical in the present study.…”

Section: Discussionsupporting

confidence: 84%

“…Molecular methods have been considered as an alternative way to identify M. pneumoniae as well as its resistance-associated mutations (conventional methods cannot evaluate drug resistance). Some molecular methods have been developed [14][15][16] to detect both M. pneumoniae and its drug resistance; however, information about an automated method is limited [17].…”

Section: Introductionmentioning

confidence: 99%

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Evaluation of GENECUBE Mycoplasma for the detection of macrolide-resistant Mycoplasma pneumoniae

Morinaga

Suzuki

Notake

et al. 2020

Journal of Medical Microbiology

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Introduction. Resistance against macrolide antibiotics in Mycoplasma pneumoniae is becoming non-negligible in terms of both appropriate therapy and diagnostic stewardship. Molecular methods have attractive features for the identification of Mycoplasma pneumoniae as well as its resistance-associated mutations of 23S ribosomal RNA (rRNA). Hypothesis/Gap Statement. The automated molecular diagnostic sytem can identify macrolide-resistant M. pneumoniae . Aim. To assess the performance of an automated molecular diagnostic system, GENECUBE Mycoplasma, in the detection of macrolide resistance-associated mutations. Methodology. To evaluate whether the system can distinguish mutant from wild-type 23S rRNA, synthetic oligonucleotides mimicking known mutations (high-level macrolide resistance, mutation in positions 2063 and 2064; low-level macrolide resistance, mutation in position 2067) were assayed. To evaluate clinical oropharyngeal samples, purified nucleic acids were obtained from M. pneumoniae -positive samples by using the GENECUBE system from nine hospitals. After confirmation by re-evaluation of M. pneumoniae positivity, Sanger-based sequencing of 23S rRNA and mutant typing using GENECUBE Mycoplasma were performed. Results. The system reproducibly identified all synthetic oligonucleotides associated with high-level macrolide resistance. Detection errors were only observed for A2067G (in 2 of the 10 measurements). The point mutation in 23S rRNA was detected in 67 (26.9 %) of 249 confirmed M. pneumoniae -positive clinical samples. The mutations at positions 2063, 2064 and 2617 were observed in 65 (97.0 %), 2 (3.0 %) and 0 (0.0 %) of the 67 samples, respectively. The mutations at positions 2063 and 2064 were A2063G and A2064G, respectively. The results from mutant typing using GENECUBE Mycoplasma were in full agreement with the results from sequence-based typing. Conclusion. GENECUBE Mycoplasma is a reliable test for the identification of clinically significant macrolide-resistant M. pneumoniae .

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Section: Discussionsupporting

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Evaluation of GENECUBE Mycoplasma for the detection of macrolide-resistant Mycoplasma pneumoniae

Morinaga

Suzuki

Notake

et al. 2020

Journal of Medical Microbiology

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“…The diagnostic method of choice for mycoplasma pneumonia is nucleic acid amplification tests like PCR and multiplex assays because they have high sensitivity and specificity compared to serologies and culture 15‐17 . Serological tests can be used when molecular tests are not available or as an adjunct to the molecular tests 18 .…”

Section: Discussionmentioning

confidence: 99%

Presenting characteristics, comorbidities, and outcomes of patients coinfected with COVID‐19 and Mycoplasma pneumoniae in the USA

Gayam

Konala

Naramala

et al. 2020

Journal of Medical Virology

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Coronavirus disease 2019 (COVID‐19) caused by severe acute respiratory syndrome‐coronavirus 2 (SARS‐CoV‐2) is spreading at a rapid pace, and the World Health Organization declared it as pandemic on 11 March 2020. Mycoplasma pneumoniae is an "atypical" bacterial pathogen commonly known to cause respiratory illness in humans. The coinfection from SARS‐CoV‐2 and mycoplasma pneumonia is rarely reported in the literature to the best of our knowledge. We present a study in which 6 of 350 patients confirmed with COVID‐19 were also diagnosed with M. pneumoniae infection. In this study, we described the clinical characteristics of patients with coinfection. Common symptoms at the onset of illness included fever (six [100%] patients); five (83.3%) patients had a cough, shortness of breath, and fatigue. The other symptoms were myalgia (66.6%), gastrointestinal symptoms (33.3%‐50%), and altered mental status (16.7%). The laboratory parameters include lymphopenia, elevated erythrocyte sedimentation rate, C‐reactive protein, lactate dehydrogenase, interleukin‐6, serum ferritin, and D‐dimer in all six (100%) patients. The chest X‐ray at presentation showed bilateral infiltrates in all the patients (100%). We also described electrocardiogram findings, complications, and treatment during hospitalization in detail. One patient died during the hospital course.

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“…In the GENECUBE system, purification mode, amplification mode or both modes can be selected for each assay; amplification mode is used for PCR of purified samples or direct PCR of prepared samples. GENECUBE is used for Mycobacterium tuberculosis [16], Mycobacterium avium, Mycobacterium intracellulare, Neisseria gonorrhoeae [17], Chlamydia trachomatis, and Mycoplasma pneumoniae [18,19]. In addition, assays for the determination of Staphylococcus aureus and mecA were released [20] and rapid precise molecular identification of the causative pathogens from positive blood culture medium without a purification process was reported.…”

Section: Introductionmentioning

confidence: 99%

Clinical evaluation of a non-purified direct molecular assay for the detection of Clostridioides difficile toxin genes in stool specimens

et al. 2020

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Recently, a new rapid assay for the detection of tcdB gene of Clostridioides difficile was developed using the GENECUBE. The assay can directly detect the tcdB gene from stool samples without a purification in approximately 35 minutes with a few minutes of preparation process. We performed a prospective comparative study of the performance of the assay at eight institutions in Japan. Fresh residual stool samples (Bristol stool scale �5) were used and comparisons were performed with the BD MAX Cdiff assay and toxigenic cultures. For the evaluation of 383 stool samples compared with the BD MAX Cdiff assay, the sensitivity, and specificity of the two assays was 99.0% (379/383), 98.1% (52/53), 99.1% (327/330), respectively. In the comparison with toxigenic culture, the total, sensitivity, and specificity were 96.6% (370/383), 85.0% (51/60), and 98.8% (319/323), respectively. The current investigation indicated the GENECUBE Clostridioides difficile assay has equivalent performance with the BD MAX Cdiff assay for the detection of tcdB gene of C. difficile.

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Rapid Detection of the Macrolide Sensitivity of Pneumonia-Causing Mycoplasma pneumoniae Using Quenching Probe Polymerase Chain Reaction (GENECUBE®)

Cited by 13 publications

References 36 publications

Evaluation of GENECUBE Mycoplasma for the detection of macrolide-resistant Mycoplasma pneumoniae

Evaluation of GENECUBE Mycoplasma for the detection of macrolide-resistant Mycoplasma pneumoniae

Presenting characteristics, comorbidities, and outcomes of patients coinfected with COVID‐19 and Mycoplasma pneumoniae in the USA

Clinical evaluation of a non-purified direct molecular assay for the detection of Clostridioides difficile toxin genes in stool specimens

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