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HelpTest Code RPMPM Mycoplasma (Mycoplasmoides) pneumoniae Macrolide (Azithromycin) Resistance Prediction, Molecular Detection, PCR, Varies
Ordering Guidance
This test should only be ordered on specimens that have tested positive for Mycoplasma (Mycoplasmoides) pneumoniae. This assay predicts M pneumoniae macrolide (Azithromycin) resistance only.
For detection of M pneumoniae prior to macrolide resistance testing , order MPRP / Mycoplasma (Mycoplasmoides) pneumoniae with Macrolide Resistance Reflex, Molecular Detection, PCR, Varies.
Necessary Information
Specimen source is required; include the specific anatomic source.
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
Secondary ID
610248Useful For
Predicting macrolide susceptibility in Mycoplasma (Mycoplasmoides) pneumoniae
Method Name
Rapid Polymerase Chain Reaction (PCR) using Light Cycler and Fluorescent Resonance Energy Transfer (FRET)
Specimen Type
VariesSpecimen Minimum Volume
Respiratory: 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 viral transport medium (VTM) or placed on a swab and then into VTM (M4-RT, M4, or M5) Body fluid specimens placed in viral transport medium (VTM) or placed on a swab and then in VTM (M4-RT, M4, or M5) |
Reject |
Reference Values
Not Predicted
Interpretation
A macrolide resistance predicted or not predicted result indicates the presence of Mycoplasma (Mycoplasmoides) pneumoniae 23S ribosomal RNA (rRNA) gene and indicates whether one of the 2 most common 23S rRNA gene single nucleotide variations (A2064G and A2063G) associated with high-level macrolide resistance is predicted.
An "unable to assess" result for M pneumoniae macrolide resistance indicates the absence of detectable M pneumoniae 23S rRNA DNA but does not negate the presence of the organism and may occur due to inhibition of the polymerase chain reaction, sequence variability underlying primers or probes, or the presence of M pneumoniae 23S rRNA DNA in quantities less than the limit of detection of the assay.
Method Description
When Mycoplasma (Mycoplasmoides) pneumoniae is detected via previous nucleic acid amplification test, a reflexive polymerase chain reaction (PCR) is performed to assess the 23S ribosomal RNA (rRNA) 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. Note, samples deemed positive outside of Mayo Clinic Laboratories may be processed according to specimen type prior to PCR testing. This includes extraction by the MagNA Pure 96 automated instrument (Roche Applied Science).
A specific target sequence of the 23S rRNA gene from M 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 PCR cycling. Detection and prediction of the M pneumoniae target is performed through melting curve analysis using the LightCycler software. While the wild-type 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. PCR inhibition is monitored through use of a recovery template.(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; 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)
Day(s) Performed
Monday through Sunday
Report Available
3 to 4 daysSpecimen Retention Time
7 daysPerforming Laboratory
Mayo Clinic Laboratories in Rochester
CPT Code Information
87798
LOINC Code Information
| Test ID | Test Order Name | Order LOINC Value |
|---|---|---|
| RPMPM | M. pneumoniae Macrolide Resist PCR | 6483-2 |
| Result ID | Test Result Name | Result LOINC Value |
|---|---|---|
| SRCMP | Specimen source | 31208-2 |
| 619928 | M. pneumoniae Macrolide Resistance | 6483-2 |
Forms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.