Mycoplasma pneumoniae – Respiratory PCR

Consistent with O. Reg. 671/92 of the French Language Services Act, laboratory testing information on this page is only available in English because it is scientific or technical in nature and is for use only by qualified health care providers and not by members of the public.

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.

*Uncontrolled print copy. Valid only on day of print: 17 July 2025.

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

General Test Requisition

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

Complete all fields of the requisition form, including:

Test(s) requests and indications for testing
Patient setting/population/source
Symptoms
Onset date of symptoms

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.

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.

*Uncontrolled print copy. Valid only on day of print: 17 July 2025.

Contact Laboratory Customer Service

Microbiology and Laboratory Services

customerservicecentre@oahpp.ca

Updated 20 May 2025