Evaluation of Five Real-Time PCR Assays for Detection of Mycoplasma pneumoniae

Dumke, Roger; Jacobs, Enno

doi:10.1128/jcm.02048-14

Cited by 18 publications

(14 citation statements)

References 21 publications

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“…Mycoplasma pneumoniae infections have been described for more than 70 years [18]. The implementation of M. pneumoniae nucleic acid amplification techniques enabled better sensitivity and specificity, and quicker results [19,20]. In this study we investigated M. pneumoniae infections resulting in ICU admission of patients in a single institution.…”

Section: Discussionmentioning

confidence: 99%

Increased rates of intensive care unit admission in patients with Mycoplasma pneumoniae : a retrospective study

Khoury

Sviri

Rmeileh

et al. 2016

Clinical Microbiology and Infection

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Mycoplasma pneumoniae is a leading cause of respiratory disease. In the Intensive Care Unit (ICU) setting M. pneumoniae is not considered a common pathogen. In 2010-13 an epidemic of M. pneumoniae-associated infections was reported and we observed an increase of M. pneumoniae patients admitted to ICU. We analysed the cohort of all M. pneumoniae-positive patients' admissions during 2007 to 2012 at the Hadassah-Hebrew University Medical Centre (a 1100-bed tertiary medical centre). Mycoplasma pneumoniae diagnosis was made routinely using PCR on throat swabs and other respiratory samples. Clinical parameters were retrospectively extracted. We identified 416 M. pneumoniae-infected patients; of which 68 (16.3%) were admitted to ICU. Of these, 48% (173/416) were paediatric patients with ICU admission rate of 4.6% (8/173). In the 19- to 65-year age group ICU admission rate rose to 18% (32/171), and to 38.8% (28/72) for patients older than 65 years. The mean APACHE II score on ICU admission was 20, with a median ICU stay of 7 days, and median hospital stay of 11.5 days. Of the ICU-admitted patients, 54.4% (37/68) were mechanically ventilated upon ICU admission. In 38.2% (26/68), additional pathogens were identified mostly later as secondary pathogens. A concomitant cardiac manifestation occurred in up to 36.8% (25/68) of patients. The in-hospital mortality was 29.4% (20/68) and correlated with APACHE II score. Contrary to previous reports, a substantial proportion (16.3%) of our M. pneumoniae-infected patients required ICU admission, especially in the adult population, with significant morbidity and mortality.

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Section: Discussionmentioning

confidence: 99%

Increased rates of intensive care unit admission in patients with Mycoplasma pneumoniae : a retrospective study

Khoury

Sviri

Rmeileh

et al. 2016

Clinical Microbiology and Infection

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“…pneumoniae and its gene is an attractive target in the clinical detection of M . pneumoniae by real-time PCR 10 17 . This is the first study detecting P1 antigen to confirm M. pneumoniae infection in children with pneumonia.…”

Section: Discussionmentioning

confidence: 99%

“…However, M. pneumoniae culture and serological tests are insensitive, time-consuming and cross-reactive; therefore, they are not appropriate for rapid and accurate detection of M. pneumoniae infection in clinical practice 9 . Recently, real-time PCR has been reported by many authors as a rapid, sensitive, and specific method, but it still requires at least 2–4 hours for DNA extraction and amplification 10 11 12 13 . In addition, real-time PCR may not discriminate live M. pneumoniae from dead ones 9 .…”

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confidence: 99%

Rapid diagnosis of Mycoplasma pneumoniae in children with pneumonia by an immuno-chromatographic antigen assay

Liu

Yun

et al. 2015

Sci Rep

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Mycoplasma pneumoniae is a particularly important pathogen that causes community acquired pneumonia in children. In this study, a rapid test was developed to diagnose M. pneumoniae by using a colloidal gold-based immuno-chromatographic assay which targets a region of the P1 gene. 302 specimens were analyzed by the colloidal gold assay in parallel with real-time PCR. Interestingly, the colloidal gold assay allowed M. pneumoniae identification, with a detection limit of 1 × 103 copies/ml. 76 samples were found to be positive in both real-time PCR and the colloidal gold assay; two specimens positive in real-time PCR were negative in the rapid colloidal gold assay. The specificity and sensitivity of the colloidal gold assay were 100% and 97.4%, respectively. These findings indicate that the newly developed immuno-chromatographic antigen assay is a rapid, sensitive and specific method for identifying M. pneumoniae, with potential clinical application in the early diagnosis of Mycoplasma pneumoniae infection.

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“…In recent years, nucleic acid-based assays targeting M. pneumoniae and C. pneumoniae have been developed (Dumke and Jacobs 2009 , 2014 ; Winchell et al 2008 ). PCR-based methods have high sensitivity, short turnaround time and the testing can be completed in a single working day.…”

Section: Introductionmentioning

confidence: 99%

Development of a multiplex real-time PCR assay for detection of Mycoplasma pneumoniae, Chlamydia pneumoniae and mutations associated with macrolide resistance in Mycoplasma pneumoniae from respiratory clinical specimens

2015

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The aim of this study was to improve detection of Mycoplasma pneumoniae and Chlamydia pneumoniae in clinical specimens by developing a multiplex real-time PCR assay that includes identification of macrolide-resistant M. pneumoniae. Novel assays targeting a M. pneumoniae conserved hypothetical protein gene, M. pneumoniae 23S rRNA gene mutations associated with macrolide resistance and human β-globin gene (an endogenous internal control) were designed and combined with a previously published C. pneumoniae PCR targeting ompA gene. The resulting quadraplex PCR was validated with a panel of clinical specimens supplemented with external quality assessment specimens, simulated specimens and various bacterial and viral strains. The obtained results were compared to those obtained by reference PCRs or confirmed by sequencing (typing of macrolide resistance). The novel multiplex PCR assay was in 100 % agreement with reference PCRs. Four M. pneumoniae strains with macrolide resistance-associated mutations were identified among 42 strains, which comprises 9.5 % of the study material. Amplification of an internal control excluded sample-derived inhibition possibly leading to false-negative reporting. In conclusion, we have developed a resources conserving multiplex real-time PCR assay for simultaneous detection of M. pneumoniae, C. pneumoniae and the most common mutations leading to macrolide resistance in M. pneumoniae. The assay is a widely useful tool for detection of these respiratory pathogens and will also shed light on the occurrence of macrolide resistance in M. pneumoniae.Electronic supplementary materialThe online version of this article (doi:10.1186/s40064-015-1457-x) contains supplementary material, which is available to authorized users.

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Evaluation of Five Real-Time PCR Assays for Detection of Mycoplasma pneumoniae

Cited by 18 publications

References 21 publications

Increased rates of intensive care unit admission in patients with Mycoplasma pneumoniae : a retrospective study

Increased rates of intensive care unit admission in patients with Mycoplasma pneumoniae : a retrospective study

Rapid diagnosis of Mycoplasma pneumoniae in children with pneumonia by an immuno-chromatographic antigen assay

Development of a multiplex real-time PCR assay for detection of Mycoplasma pneumoniae, Chlamydia pneumoniae and mutations associated with macrolide resistance in Mycoplasma pneumoniae from respiratory clinical specimens

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