Mycoplasma pneumoniae – Respiratory PCR

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Background
This page provides real-time PCR testing information for Mycoplasma pneumoniae at Public Health Ontario (PHO).

Updates
Cerebrospinal Fluid (CSF) is added under specimen type. Prior to submission of CSF to Public Health Ontario, contact PHO’s Laboratory Customer Service Centre for Microbiologist’s approval to avoid rejection or delay in testing.

Testing Indications

Mycoplasma pneumoniae is a common cause of upper respiratory tract infections, bronchitis, and “atypical” community-acquired pneumonia. Clinical manifestations outside the respiratory tract are rare but may occur. M. pneumoniae is transmitted via respiratory droplets and can cause sporadic infections and sustained outbreaks.

If an outbreak is suspected, contact your local Public Health Unit.

Acceptance/Rejection Criteria

Acceptable:

  • Use recommended collection kit (see below).
  • Submit neat specimen (>=0.5 ml)
  • Swab specimens must be collected with nylon flocculated swabs or on swabs with Dacron tips and aluminum or plastic shafts.

Unacceptable:

  • Swabs with calcium alginate or cotton tips and wooden sticks are not acceptable, as they have been shown to inhibit PCR.
  • Chemically fixed or preserved specimens are not acceptable for PCR testing and will be rejected upon receipt at PHO.
  • Cobas PCR media is unacceptable as a transport media for Mycoplasma pneumoniae.  Specimens sent in this media will be rejected.

Specimen Collection and Handling

For outbreaks contact Medical/Clinical Microbiologist at PHO (Customer Service Center 1-877 604 4567).

Specimen Requirements

Test Requested Required Requisition(s) Specimen Type Minimum Volume Collection Kit

Mycoplasma pneumoniae PCR

Bronchoalveolar lavage

Nasopharyngeal Swab (NPS)

Throat swab

Tracheal aspirates

Respiratory tract specimens

Cerebrospinal Fluid(see note 3)

- Swab in universal transport media provided in collection  kit

- Neat (minimum volume 0.5mL)

Mycoplasma pneumoniae / Chlamydia pneumoniae KIT order# 390085 (6 Packs).

Submission and Collection Notes

1

Complete all fields of the requisition form, including:

  1. Test(s) requests and indications for testing
  2. Patient setting/population/source
  3. Symptoms
  4. Onset date of symptoms
2

For clinical specimens, label the specimen container(s) with the patient’s first and last name, date of collection, and one other unique identifier such as the patient’s date of birth or Health Card Number. For additional information see: Criteria for Acceptance of Patient Specimens. Failure to provide this information may result in rejection or testing delay.

3

Before submitting CSF specimen to PHO, contact PHO’s Laboratory Customer Service Centre first for a Microbiologist’s approval. Note that CSF is not a validated specimen type.

Timing of Specimen Collection

Specimens should be collected as soon as possible following onset of symptoms using the Mycoplasma pneumoniae / Chlamydophila pneumoniae collection kit.

Storage and Transport

Specimens placed inside a biohazard bag and sealed should be stored at 2-8°C following collection and shipped to Public Health Ontario on ice packs. All clinical specimens must be shipped in accordance to the Transportation of Dangerous Good Act.

Requisitions and Kit Ordering

Test Frequency and Turnaround Time (TAT)

Mycoplasma pneumoniae PCR testing is performed at PHO, Toronto and Kingston locations.

TAT is up to 5 business days from receipt at Public Health Ontario.

Stat and Critical Specimens Testing

Contact PHO’s Laboratory Customer Service Centre when submitting critical specimen so it will be prioritized and included in the next available testing run.

Test Methods

The method is a laboratory-developed multiplex real- polymerase chain reaction (PCR)which detects Community-Acquired Respiratory Distress Syndrome (CARDS) toxin gene (Thurman et. al.) in Mycoplasma pneumoniae.

Only respiratory tract specimens like bronchoalveolar lavage (BAL), nasopharyngeal swab (NPS), throat swab, and tracheal aspirates were validated for this assay at PHO. Cerebrospinal fluid (CSF) is not a validated specimen type but can be tested when approved by the PHO microbiologist.

Interpretation

The following table provides possible test results with associated interpretations:

Result Interpretation Comments

Not Detected

Mycoplasma pneumoniae Not Detected

Note: This specimen was tested using a laboratory test developed by Public Health Ontario Laboratory for the purpose of clinical testing. Results should be interpreted in the context of the clinical situation.

Detected

Mycoplasma pneumoniae Detected

Note: This specimen was tested using a laboratory test developed by Public Health Ontario Laboratory for the purpose of clinical testing. Results should be interpreted in the context of the clinical situation.

Invalid

Mycoplasma pneumoniae Invalid

Note: Nucleic Acid extraction and/or amplification could not be successfully completed.  This may be caused by inhibitory substances or improper sample collection.  Please resubmit if clinically relevant.

This specimen was tested using a laboratory test developed by Public Health Ontario Laboratory for the purpose of clinical testing. Results should be interpreted in the context of the clinical situation.

Indeterminate

Mycoplasma pneumoniae Indeterminate

Note: An indeterminate PCR test result may be due to a low level of target genetic material in the specimen content or a non-specific signal.  Please resubmit another specimen for testing if clinically indicated.

This specimen was tested using a laboratory test developed by Public Health Ontario Laboratory for the purpose of clinical testing. Results should be interpreted in the context of the clinical situation.

Inconclusive

Mycoplasma pneumoniae Inconclusive

Note: PCR results are inconclusive.  Consider collecting a second specimen for repeat PCR testing.

This specimen was tested using a laboratory test developed by Public Health Ontario Laboratory for the purpose of clinical testing. Results should be interpreted in context of the clinical situation.

Reporting

Results are reported to the physician, authorized health care provider (General O. Reg 45/22, s.18) or submitter as indicated on the requisition.

Mis à jour le 20 mai 2025