Diagnosis of infections due to Mycoplasmoides pneumoniae
Rapid Polymerase Chain Reaction (PCR) using Light Cycler and Fluorescent Resonance Energy Transfer (FRET)
Mycoplasma pneumoniae
Mycoplasmoides pneumoniae
Varies
Specimen source is required.
Question ID | Description | Answers |
---|---|---|
SRCMP | Specimen Source |
The high sensitivity of amplification by polymerase chain reaction requires the specimen to be processed in an environment in which contamination of the specimen by Mycoplasmoides pneumoniae DNA is unlikely.
Submit only 1 of the following specimens:
Specimen Type: Respiratory
Supplies:
Sources: Bronchial washing, bronchoalveolar lavage, tracheal secretions, sputum
Container/Tube:
Acceptable: Specimen in M4, M4-RT, M5, M6, or universal transport medium
Specimen Volume: 1 mL
Specimen Type: Swab
Supplies:
-Culturette (BBL Culture Swab) (T092)
-BD Eswab (T853)
-Culture Swab-Liquid Stuarts/Single Swab (NP Swab) (T515)
-Nasopharyngeal Swab (Nylon Mini-Tip Swab) (T861)
-M4-RT (T605)
Sources: Throat, nasal, or nasopharyngeal
Container/Tube:
Acceptable: Culture transport swab (Stuart's media) or place swab in M4, M4-RT, M5, M6, universal transport media, or ESwab
Specimen Volume: Swab
Collection Instructions:
1. Collect specimen by swabbing back and forth over mucosa surface to maximize recovery of cells.
2. Place swab back into swab cylinder.
Specimen Type: Fluid
Sources: Pleural, pericardial, cerebrospinal
Container/Tube: Sterile vial
Specimen Volume: 0.5 mL
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.
Respiratory, Fluid: 0.5 mL
Swab: 1 swab
Cotton or calcium alginate-tipped swab, wooden shaft swab, transport swab containing gel or charcoal Port-a-Cul tube Anaerobic fluid vials Dry swab (no pledget or sponge) | Reject |
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Varies | Refrigerated (preferred) | 7 days | |
Frozen | 7 days |
Diagnosis of infections due to Mycoplasmoides pneumoniae
Mycoplasmoides pneumoniae, previously Mycoplasma pneumoniae, is a small bacterium transmitted via organism-containing droplets. It is a cause of upper respiratory infection, pharyngitis, and tracheobronchitis, particularly in children, and has been associated with approximately 20% of cases of community acquired pneumonia.(1) Central nervous system and cardiac manifestations are probably the most frequent extrapulmonary complications of infections due to M pneumoniae. The disease is usually self-limited although severe disease has been reported in immunocompromised patients.(2)
Identification of M pneumoniae by culture-based methods is time consuming and insensitive. Serology based assays for M pneumoniae have several drawbacks. The development of IgM antibodies takes approximately 1 week and the IgM response in adults may be variable or it may be decreased in immunosuppressed individuals.(3,4) Confirmation of the disease may be dependent on the observation of a 4-fold rise in IgG antibody titers between acute and convalescent specimens, several weeks following the initial onset of illness, providing clinical utility only for retrospective testing.(4) Real-time polymerase chain reaction testing offers a rapid and sensitive option for detection of M pneumoniae DNA from clinical specimens.
Not applicable
A positive result indicates the presence of Mycoplasmoides pneumoniae.
A negative result does not rule out the presence of M pneumoniae and may be due to the presence of inhibitors within the specimen matrix, or the presence of organisms at numbers below the limits of detection of the assay.
This assay should only be used for testing of respiratory tract specimens (throat swabs, nasopharyngeal swabs, tracheal secretions, sputum, and bronchoalveolar lavage fluid) and pleural/chest fluid, pericardial fluid, and cerebrospinal fluid.
Accuracy:
The assay was validated in a blinded manner using 30 Mycoplasma pneumoniae-positive specimens received from a reference lab and 6 negative specimens. The M pneumoniae polymerase chain reaction (PCR) test had 100% sensitivity and specificity when compared to the Focus Diagnostics M pneumoniae primer pair PCR assay. Whole organism spiking studies (near the limit of detection of the assay) were also performed using the following specimens: bronchoalveolar lavage/bronchial wash, nasopharyngeal and throat swabs, sputum, pericardial/pleural fluid, and cerebrospinal fluid. These specimens were confirmed as being negative for M pneumoniae prior to spiking. The sensitivity and specificity of the spiked specimens combined for all the matrices were 99% (154/155) and 100% (57/57), respectively.
Limit of detection:
The limit of detection of the assay is less than 5 target copies/mcL for all validated specimen types.
Analytical specificity:
The assay was tested against a panel of 45 organisms consisting of bacteria and viruses representing normal respiratory flora and/or respiratory pathogens. There was no cross reactivity among these organisms, which included 16 other species of Mycoplasmoides.
3. Daxboeck F, Krause R, Wenisch C: Laboratory diagnosis of Mycoplasma pneumoniae infection. Clin Microbiol Infect. 2003;9:263-273
Throat swabs, nasopharyngeal swabs, sputum, bronchoalveolar lavage fluid, pericardial/pleural/chest fluid, and cerebrospinal fluid specimens are processed according to specimen type. Nucleic acid is extracted by the MagNA Pure automated instrument (Roche Applied Science). A specific target sequence from Mycoplasma pneumoniae is targeted by primers and fluorescence resonance energy transfer (FRET) hybridization probes. The LightCycler instrument (Roche Applied Science) amplifies and monitors the development of target nucleic acid sequences after the annealing step during polymerase chain reaction cycling. Detection of the M pneumoniae target is performed through melting curve analysis using the LightCycler software.(Schmitt BH, Sloan LM, Patel R: Real-time PCR detection of Mycoplasma pneumoniae in respiratory specimens. Diagn Microbiol Infect Dis. 2013 Nov;77[3]:202-205)
Monday through Sunday
This test was developed, and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. This test has not been cleared or approved by the US Food and Drug Administration.
87581
Test Id | Test Order Name | Order LOINC Value |
---|---|---|
MPRP | Mycoplasma pneumoniae PCR | 29257-3 |
Result Id | Test Result Name |
Result LOINC Value
Applies only to results expressed in units of measure originally reported by the performing laboratory. These values do not apply to results that are converted to other units of measure.
|
---|---|---|
SRCMP | Specimen source | 31208-2 |
62394 | Mycoplasma pneumoniae PCR | 29257-3 |