Efficiency of the novel quenching-probe PCR method to detect 23S rRNA mutations in children with Mycoplasma pneumoniae infection

Nagita, Akira; Muramatsu, Hirotada; Hokama, Madoka; Takami, Masao; Murakami, Yoshihiro; Funashima, Yumiko; Nagasawa, Zenzo

doi:10.1016/j.mimet.2021.106135

Cited by 7 publications

(10 citation statements)

References 14 publications

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“…Reagents for the diagnosing of Mycoplasma pneumoniae and coronavirus disease 2019 infections are already on the market in Japan and are widely used in clinical settings. [13][14][15] The reagents developed by us for H. pylori genes and the CAM-resistant mutation had comparable accuracy with real-time PCR and sequencing analysis. Therefore, our findings suggest that it could be used in daily medical care.…”

Section: Discussionmentioning

confidence: 97%

“…It is compact, inexpensive, and does not require any special procedure; therefore, it can be set in clinics as just a point‐of‐care testing device for PCR. Reagents for the diagnosing of Mycoplasma pneumoniae and coronavirus disease 2019 infections are already on the market in Japan and are widely used in clinical settings 13–15 . The reagents developed by us for H. pylori genes and the CAM‐resistant mutation had comparable accuracy with real‐time PCR and sequencing analysis.…”

Section: Discussionmentioning

confidence: 99%

“…At 8-12 weeks after the eradication therapy, the success of therapy was assessed by a 13 C-urea breath test. Breath samples were obtained 4 h after a meal and 20 min after ingestion of 100-mg 13 C-urea (UBIT ™ tablets 100 mg).…”

Section: Sequencing Analysis (Sanger Method)mentioning

confidence: 99%

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Smart Gene™ as an effective non‐invasive point‐of‐care test to detect Helicobacter pylori clarithromycin‐resistant mutation

Kakiuchi

Okuda

Matsuo

et al. 2022

J of Gastro and Hepatol

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Background and Aim: Smart Gene ™ was developed based on the concept of point-ofcare genetic testing. We evaluated the detection performance of a reagent for Helicobacter pylori (H. pylori) clarithromycin (CAM)-resistant mutation assessment and determined the association between the results of Smart Gene ™ and those of eradication therapy for H. pylori. Methods: In 2020, the present study was conducted on participants of the H. pylori test and treat project in Saga Prefecture. The submitted stool samples were measured for H. pylori gene and CAM-resistant mutation by Smart Gene ™ , and the results were compared with real-time polymerase chain reaction (PCR) and sequencing analysis. Finally, the results of the eradication therapy were examined for each result of Smart Gene ™ . Results: Stool samples were obtained from 139 patients who were tested positive by stool antigen test and were analyzed. The H. pylori detection rate was 95.7% by Smart Gene ™ , 92.8% by real-time PCR (P < 0.01), and 89.2% by sequencing analysis (P = 0.06). The overall concordance rate for CAM-resistant mutation between Smart Gene ™ and sequencing analysis was 96.7%. Moreover, 35 of 48 students with CAM-resistant mutation and 33 of the 35 students with a mutation without CAM resistance succeeded in CAM-containing triple therapy, and the success rate was significantly higher for the mutation without CAM resistance (P = 0.012). Conclusions:The detection performance of Smart Gene ™ was comparable with that of real-time PCR and sequencing analysis. It is expected that the success rate of eradication would be further improved by using the reagent.

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

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Smart Gene™ as an effective non‐invasive point‐of‐care test to detect Helicobacter pylori clarithromycin‐resistant mutation

Kakiuchi

Okuda

Matsuo

et al. 2022

J of Gastro and Hepatol

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“…pneumoniae were detected in 24 of 38 patients infected with M . pneumoniae using the Smart Gene ® system and the results were confirmed by direct sequencing, revealing all mutations as A2063G [ 61 ].…”

Section: Discussionmentioning

confidence: 99%

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

Ishiguro

Sato

Mori³

et al. 2021

PLoS ONE

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Objectives 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® 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® system for detection of M. pneumoniae and detection of a point mutation at domain V of the 23S rRNA gene of M. pneumoniae. Materials Pharyngeal swab samples were collected from 154 patients who were suspected of having respiratory tract infections associated with M. pneumoniae. Results Compared with the results of qPCR, the sensitivity and specificity of the Smart Gene® 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® 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 μg/ml and those among the 33 isolates without a mutation in the 23S rRNA gene were <0.0625 μg/ml. Conclusion The Smart Gene® 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® system is suitable for point-of-care testing in both hospital and outpatient settings.

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“…Smart Gene™ (MIZUHO MEDY Co., Ltd.) was developed on the concept of point‐of‐care testing (POCT), which can automatically perform nucleic acid extraction, amplification, and detection. The detection of Mycoplasma pneumoniae and coronavirus by Smart Gene™ with pharyngeal swabs shows high reproducibility and useability and allows quick patient triage 8–11 . In addition, the detection of H. pylori by Smart Gene™ with stool proved to be an effective non‐invasive test 12 .…”

Section: Introductionmentioning

confidence: 99%

Clinical evaluation of a novel molecular diagnosis kit for detecting Helicobacter pylori and clarithromycin‐resistant using intragastric fluid

et al. 2022

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Background: Although there are many Helicobacter pylori (H. pylori) diagnostic methods, the culture and antibiotic susceptibility test is an important method for selecting the most effective H. pylori eradication regimen. However, this diagnostic method is complicated and takes several days; therefore, the development of a rapid and simple diagnostic method is required. Eradication failure due to clarithromycin (CAM) resistance should also be considered. In this study, we report the clinical evaluation of point-of-care testing (POCT) kit using intragastric fluid, a novel kit for detecting H. pylori and CAM resistance. Materials and Methods:The study participants were 143 patients suspected of H. pylori infection and had an endoscopic examination. The novel diagnostic kit diagnosed H. pylori infection and CAM resistance-associated mutation using intragastric fluid. To diagnose H. pylori infection, the relationship between the diagnostic kit and conventional diagnostic methods (urea breath test, stool antigen test, culture test, and realtime polymerase chain reaction [PCR]) was evaluated. For CAM resistance-associated mutation detection, the concordance between the diagnostic kit and antibiotic susceptibility test was evaluated. Results:The diagnosis of H. pylori infection with the novel molecular diagnostic kit using intragastric fluid showed significant relationship with conventional diagnostic methods. Especially when the culture was control, the sensitivity was 100% (67/67), the specificity was 95.9% (71/74), and the overall concordance was 97.9% (138/141).The detection of CAM resistance-associated mutations had a concordance rate of 97.0% (65/67) when compared with the antibiotic susceptibility test. Conclusions:The H. pylori molecular POCT kit uses intragastric fluid as a sample and can diagnose H. pylori infection and detect CAM resistance-associated mutations within an hour. This novel kit is expected to prove useful in selecting the most effective eradication regimen for H. pylori.

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Efficiency of the novel quenching-probe PCR method to detect 23S rRNA mutations in children with Mycoplasma pneumoniae infection

Cited by 7 publications

References 14 publications

Smart Gene™ as an effective non‐invasive point‐of‐care test to detect Helicobacter pylori clarithromycin‐resistant mutation

Smart Gene™ as an effective non‐invasive point‐of‐care test to detect Helicobacter pylori clarithromycin‐resistant mutation

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

Clinical evaluation of a novel molecular diagnosis kit for detecting Helicobacter pylori and clarithromycin‐resistant using intragastric fluid

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