ABSTRACT
BACKGROUND: Patients with acute respiratory tract infections are frequently prescribed antimicrobials despite high rates of virus detection. Physicians may overprescribe antimicrobials owing to the concern of bacterial infections, including those because of atypical pathogens. We investigated the accuracy of clinical predictions concerning atypical pathogen infections. METHODS: We prospectively enrolled adult patients who presented with a fever and cough in outpatient clinics between December 2016 and August 2018. After taking a history and performing physical examinations, physicians predicted the possibility of respiratory infections because of atypical pathogens. Disease probabilities were categorized into 3 grades (high: ≥50%, intermediate: 20% ≥ and <50%, and low: <20%) and were judged by physicians who were taking care of the patients. Confirmation of atypical pathogens was performed by comprehensive molecular analyses of respiratory samples. RESULTS: Atypical pathogens were detected in 21 of 210 patients. A close contact history (odds ratio [OR]: 11.4, 95% confidence interval [CI]: 2.4-53.5) and the presence of pneumonia (OR: 12.9, CI: 4.3-39.2) were associated with the detections. Atypical pathogens were detected in 32.3% of high-probability cases (10/31), while atypical pathogens were only detected in 8.8% of intermediate-probability cases (8/91) and 3.4% of low-probability cases (3/88) (P < .001). CONCLUSIONS: The current study indicates that physicians' predictions were associated with the detection of atypical pathogens; however, overestimation was observed.
ABSTRACT
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 (26.9â%) of 249 confirmed M. pneumoniae-positive clinical samples. The mutations at positions 2063, 2064 and 2617 were observed in 65 (97.0â%), 2 (3.0â%) and 0 (0.0â%) 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.
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Macrolides/pharmacology , Molecular Diagnostic Techniques/methods , Mycoplasma pneumoniae/isolation & purification , Pneumonia, Mycoplasma/diagnosis , Automation , DNA, Bacterial , Humans , Mutation , Mycoplasma pneumoniae/drug effects , Mycoplasma pneumoniae/genetics , Oligonucleotides/chemical synthesis , Pneumonia, Mycoplasma/microbiology , Polymerase Chain Reaction/methods , Sequence Analysis, DNAABSTRACT
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.
