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Test ID MPRP Mycoplasma (Mycoplasmoides) pneumoniae with Macrolide Resistance Reflex, Molecular Detection, PCR, Varies

Useful For

Diagnosing infections due to Mycoplasma (Mycoplasmoides) pneumoniae

 

Assessing macrolide susceptibility

Reporting Name

M pneumoniae PCR + Macrolide Reflex

Specimen Type

Varies


Necessary Information


Specimen source is required.



Specimen Required


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 Mycoplasma (Mycoplasmoides) pneumoniae DNA is unlikely.

 

Submit only 1 of the following specimens:

 

Specimen Type: Swab

Supplies:

-Culturette (BBL Culture Swab) (T092)

-BD E-Swab (T853)

-Nasopharyngeal Swab (Nylon Mini-Tip Swab) (T861)

-Culture Swab-Liquid Stuarts/Single Swab (NP Swab) (T515)

-M4-RT (T605)

Sources: Throat, nasal, or nasopharyngeal

Container/Tube:

Preferred: Culture swab transport system (Dacron or rayon swab with aluminum or plastic shaft with either Stuart or Amies liquid medium)

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 container

Specimen Volume: 0.5 mL

 

Specimen Type: Respiratory

Sources: Bronchial washing, bronchoalveolar lavage, tracheal secretions, sputum

Container/Tube: Sterile container

Specimen Volume: 1 mL


Specimen Minimum Volume

Respiratory specimen: 0.5 mL
Other specimen types: See Specimen Required

Specimen Stability Information

Specimen Type Temperature Time Special Container
Varies Refrigerated (preferred) 7 days
  Frozen  7 days

Reject Due To

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)
Respiratory fluid specimens placed in VTM or placed on a swab and then into VTM (M4-RT, M4, or M5)
Body fluid specimens placed in VTM or placed on a swab and then in VTM (M4-RT, M4, or M5)		 Reject

Clinical Information

Mycoplasma (Mycoplasmoides) 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 some of the extrapulmonary complications of infections due to M pneumoniae. The disease is usually self-limited although severe disease may occur, including in patients who are immunocompromised.(2)

 

Identification of M pneumoniae by culture-based methods is time consuming and insensitive. Serologic assays have drawbacks; the development of IgM antibodies takes approximately 1 week, and the IgM response in adults may be variable or 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, only after several weeks following the initial onset of illness, only providing clinical application for retrospective testing and not individual patient care.(4) Real-time polymerase chain reaction (PCR) testing offers a rapid and sensitive option for detection of M pneumoniae DNA from clinical specimens.(5)

 

Macrolide resistance in M pneumoniae is increasingly reported. In a study performed at Mayo Clinic, 10% of M pneumoniae detections were associated with macrolide resistance.(6) Real-time PCR testing can be used to assess for common mutations associated with macrolide resistance in M pneumoniae.

Reference Values

Negative

Interpretation

A positive result indicates the presence of Mycoplasma (Mycoplasmoides) pneumoniae. If detected, common mutations associated with macrolide resistance in M pneumoniae may be assessed.

 

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 target DNA below the limit of detection of the assay.

Cautions

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.

Clinical Reference

1. Waites KB, Taylor-Robinson D: Mycoplasma and Ureaplasma. In. Versalovic J, Carroll K, Funke G, et al, eds. Manual of Clinical Microbiology. ASM Press; 2011:970-985

2. Jensen JS, Heilmann C, Valerius NH. Mycoplasma pneumoniae infection in a child with AIDS. Clin Infect Dis. 1994;19(1):207

3. Daxboeck F, Krause R, Wenisch C. Laboratory diagnosis of Mycoplasma pneumoniae infection. Clin Microbiol Infect. 2003;9(4):263-273

4. Waites KB, Talkington DF. Mycoplasma pneumoniae and its role as a human pathogen. Clin Microbiol Rev. 2004;17(4):697-728

5. Schmitt BH, Sloan LM, Patel R. Real-time PCR detection of Mycoplasma pneumoniae in respiratory specimens. Diagn Microbiol Infect Dis. 2013;77(3):202-205

6. Rothstein TE, Cunningham SA, Rieke RA, Mainella JM, Mutchler MM, Patel R. Macrolide resistance in Mycoplasma pneumoniae, Midwestern United States, 2014 to 2021. Antimicrob Agents Chemother. 2022;66(4):e0243221

Method Description

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 96 automated instrument (Roche Applied Science). A specific target sequence from Mycoplasma (Mycoplasmoides) pneumoniae is targeted by primers and fluorescence resonance energy transfer hybridization probes. The LightCycler 480 II instrument (Roche Applied Science) amplifies and monitors the development of target nucleic acid sequences after the annealing step during polymerase chain reaction (PCR) 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)

 

If M pneumoniae is detected, a reflexive PCR is performed to assess the 23S ribosomal RNA gene region of M pneumoniae and predict macrolide resistance based on the most common, high-level point mutations at positions 2064 and 2063 via melting curve analysis. While the wildtype genotype will display a stable melting temperature, the designed primer and probe combinations will be highly sensitive to single nucleotide mutations resulting in a cooler (left shift) melting temperature value.(Rothstein TE, Cunningham SA, Rieke RA, Mainella JM, Mutchler MM, Patel R. Macrolide resistance in Mycoplasma pneumoniae, Midwestern United States, 2014 to 2021. Antimicrob Agents Chemother. 2022 Apr 19;66[4]:e0243221)

Day(s) Performed

Monday through Sunday

Report Available

3 to 4 days

Test Classification

This test was developed and its performance characteristics determined by Mayo Clinic in a manner consistent with CLIA requirements. It has not been cleared or approved by the US Food and Drug Administration.

CPT Code Information

87581

NY State Approved

Yes

Method Name

Rapid Polymerase Chain Reaction (PCR) using Light Cycler and Fluorescent Resonance Energy Transfer (FRET)

Forms

If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
RPMPM M. pneumoniae Macrolide Resist PCR Yes No

Testing Algorithm

If positive, Mycoplasma pneumoniae macrolide resistance will be performed at an additional charge.