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

• Published in: PloS one Vol. 16; no. 10; p. e0258694
• Main Authors: Ishiguro, Nobuhisa, Sato, Rikako, Mori, Toshihiko, Tanaka, Hiroshi, Narita, Mitsuo, Nagano, Takashi, Owaku, Masato, Miyajima, Kensuke, Manabe, Atsushi
• Format: Journal Article
• Language: English
• Published: San Francisco Public Library of Science 14.10.2021; Public Library of Science (PLoS)
• Subjects: Acids; Analysis; Antibiotics; Antimicrobial agents; Asthma; Biology and Life Sciences; Cartilage; Clarithromycin; Diagnosis; Domains; Drug resistance; Evaluation; Funding; Hospitals; Infections; Infectious diseases; Medical diagnosis; Medicine and Health Sciences; Molecular diagnostic techniques; Mutation; Mycoplasma pneumoniae; Pathogens; Patients; Pediatrics; Pharynx; Pneumonia; Point mutation; Point of care testing; Polymerase chain reaction; Research and Analysis Methods; Respiratory tract; Respiratory tract diseases; rRNA; rRNA 23S; Sensitivity; University graduates; Japan; Asia; United States > US; Europe
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0258694

Macrolides are generally considered to be the drugs of choice for treatment of patients with Mycoplasma pneumoniae infection. However, macrolide-resistant M. pneumoniae has been emerging since about 2000. The Smart Gene.sup.® system (MIZUHO MEDY Co., Ltd., Tosu, Japan) is a novel fully automated system for detection of pathogens using the method of quantitative polymerase chain reaction (qPCR) with QProbe (QProbe PCR). The entire procedure is completed within 50 min and the size of the instrument is small (15 x 34 x 30 cm). The purpose of this study was to evaluate the usefulness of the Smart Gene.sup.® system for detection of M. pneumoniae and detection of a point mutation at domain V of the 23S rRNA gene of M. pneumoniae. Pharyngeal swab samples were collected from 154 patients who were suspected of having respiratory tract infections associated with M. pneumoniae. Compared with the results of qPCR, the sensitivity and specificity of the Smart Gene.sup.® system were 98.7% (78/79) and 100.0% (75/75), respectively. A point mutation at domain V of the 23S rRNA gene was detected from 7 (9.0%) of 78 M. pneumoniae-positive samples by the Smart Gene.sup.® system and these results were confirmed by direct sequencing. The minimum inhibitory concentrations of clarithromycin among the 5 isolates of M. pneumoniae with a point mutation at domain V of the 23S rRNA gene were >64 [mu]g/ml and those among the 33 isolates without a mutation in the 23S rRNA gene were <0.0625 [mu]g/ml. The Smart Gene.sup.® system is a rapid and accurate assay for detection of the existence of M. pneumoniae and a point mutation at domain V of the 23S rRNA gene of M. pneumoniae at the same time. The Smart Gene.sup.® system is suitable for point-of-care testing in both hospital and outpatient settings.