Clinical utility of loop-mediated isothermal amplification for rapid diagnosis of Mycoplasma pneumoniae in children

Aizawa, Yuta; Oishi, Tomohiro; Tsukano, Shinya; Taguchi, Tetsuo; Saitoh, Akihiko

doi:10.1099/jmm.0.068288-0

Cited by 17 publications

(18 citation statements)

References 18 publications

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“…Sensitivity of the IC test remained at 60% at 2 days, but increased to >80% at 3 days after the onset of fever and further rose after that. This could indicate that M. pneumoniae proliferates and bacterial count gradually increases over several days in the lung in subjects with M. pneumoniae infection, as previously described by Aizawa et al . M. pneumoniae would be transported to the upper respiratory tract with cough, and reflected by higher M. pneumoniae antigen quantity there.…”

Section: Discussionsupporting

confidence: 76%

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Immunochromatography test for rapid diagnosis of Mycoplasma pneumoniae infection

Onari¹,

Okada²,

Okada

et al. 2017

Pediatrics International

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The sensitivity and specificity of a new rapid Mycoplasma pneumoniae antigen immunochromatography (IC) test, DK-MP-001, were determined using particle agglutination (PA) antibody response and loop-mediated isothermal amplification (LAMP) gene detection as the gold standard. Of 165 patients, 59 were diagnosed with M. pneumoniae infection based on a ≥fourfold rise of serum PA antibody during the course of the illness. Of the first visit swabs, 60 were positive for M. pneumoniae on LAMP, and 49 were positive for M. pneumoniae antigen on IC test. Compared with PA antibody and LAMP, the sensitivity/specificity of the IC test were 81.4% (48/59) and 99.1% (105/106); and 81.7% (49/60) and 100% (105/105), respectively. IC test detected antigen in pharyngeal swabs more sensitively than in nasal swabs for the same subjects (P < 0.05). The IC test performs well enough to be used with pharyngeal swabs at the first examination.

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Section: Discussionsupporting

confidence: 76%

“…This suggests that proliferating M. pneumoniae in the lower respiratory tract may have been transferred with cough to the pharynx but not to the nasal cavity . These results have again confirmed the adequate timing and sampling site for efficient rapid diagnosis of M. pneumoniae infection …”

Section: Discussionmentioning

confidence: 56%

Immunochromatography test for rapid diagnosis of Mycoplasma pneumoniae infection

Onari¹,

Okada²,

Okada

et al. 2017

Pediatrics International

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“…In particular, adding HNB dye to the reaction tube before amplification has improved the capacity of the LAMP method to be used as a point-of-care diagnostic test, since it has made the assay easier to use in resource-limited settings (32,33). Recently, LAMP technology has been applied with high accuracy to detect infection with a wide array of pathogens, including bacteria (34,35), viruses (36), and parasites (28,37,38).…”

mentioning

confidence: 99%

Loop-Mediated Isothermal Amplification for Rapid and Semiquantitative Detection of Loa loa Infection

et al. 2014

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cRapid and accurate tests are currently needed to identify individuals with high levels of Loa loa microfilaria (mf), so that these individuals may be excluded from mass ivermectin administration campaigns against onchocerciasis and lymphatic filariasis being conducted in areas where Onchocerca volvulus, Wuchereria bancrofti, and L. loa are coendemic. To address this need, colorimetric loop-mediated isothermal amplification (LAMP) assays targeting the L. loa-specific gene sequences LLMF72 and LLMF342 were developed for the detection and quantification of L. loa microfilaremia. Both LAMP assays were highly specific (100%) for L. loa infection compared to the absence of infection or infection with related filarial pathogens. The LLMF72-based LAMP assay showed greater analytic sensitivity (limit of detection, 0.1 pg/ml of genomic DNA [gDNA] and/or 5 mf/ml) than the LLMF342-based LAMP assay (10 pg/ml of gDNA and/or 50 mf/ml), and its analytic sensitivity was similar to that of LLMF72-based quantitative PCR (qPCR). A high level of correlation was observed between microfilaria counts as determined by LLMF72-based qPCR and time to positivity by the LAMP assay, and performance measures of sensitivity, specificity, and positive and negative predictive values were similar for both assays when applied to field-collected clinical samples. By simply varying the run time, the LAMP assay was able to accurately distinguish individuals at risk for serious adverse events (SAEs) after exposure to ivermectin, using thresholds of >5,000 mf/ml and >30,000 mf/ml as indicators of increasing levels of risk. In summary, LLMF72 LAMP represents a new molecular diagnostic tool that is readily applicable as a point-of-care method for L. loa microfilarial detection and quantification in resource-limited countries where L. loa infection is endemic.

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“…Previous methods used to detect MPs had limitations (Daxboeck et al, 2003;Long et al, 2012;Yuta et al, 2014;Katsushima et al, 2015). In clinical practice, co-infections are common in children with pneumonia.…”

Section: Discussionmentioning

confidence: 99%

Screening and Identification of APOC1 as a Novel Potential Biomarker for Differentiate of Mycoplasma pneumoniae in Children

Sun

et al. 2016

Front. Microbiol.

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Background: Although Mycoplasma pneumoniae (MP) is a common cause of community-acquired pneumonia (CAP) in children, the currently used diagnostic methods are not optimal. Proteomics is increasingly being used to study the biomarkers of infectious diseases.Methods: Label-free quantitative proteomics and liquid chromatography-mass/mass spectrometry were used to analyze the fold change of protein expression in plasma of children with MP pneumonia (MPP), infectious disease control (IDC), and healthy control (HC) groups. Selected proteins that can distinguish MPP from HC and IDC were further validated by enzyme-linked immunosorbent assay (ELISA).Results: After multivariate analyses, 27 potential plasma biomarkers were identified to be expressed differently among child MPP, HC, and IDC groups. Among these proteins, SERPINA3, APOC1, ANXA6, KNTC1, and CFLAR were selected for ELISA verification. SERPINA3, APOC1, and CFLAR levels were significantly different among the three groups and the ratios were consistent with the trends of proteomics results. A comparison of MPP patients and HC showed APOC1 had the largest area under the curve (AUC) of 0.853, with 77.6% sensitivity and 81.1% specificity. When APOC1 levels were compared between MPP and IDC patients, it also showed a relatively high AUC of 0.882, with 77.6% sensitivity and 85.3% specificity.Conclusion: APOC1 is a potential biomarker for the rapid and noninvasive diagnosis of MPP in children. The present finding may offer new insights into the pathogenesis and biomarker selection of MPP in children.

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Clinical utility of loop-mediated isothermal amplification for rapid diagnosis of Mycoplasma pneumoniae in children

Cited by 17 publications

References 18 publications

Immunochromatography test for rapid diagnosis of Mycoplasma pneumoniae infection

Immunochromatography test for rapid diagnosis of Mycoplasma pneumoniae infection

Loop-Mediated Isothermal Amplification for Rapid and Semiquantitative Detection of Loa loa Infection

Screening and Identification of APOC1 as a Novel Potential Biomarker for Differentiate of Mycoplasma pneumoniae in Children

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