Molecular Detection by PCR

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Requisition Forms

Reference Details

Description:

Molecular detection of Mycoplasma pneumoniae by PCR.

 

Test Category:
Molecular Detection
Pathogen:
Mycoplasma pneumoniae
Illnesses and Diseases:
  • Atypical pneumonia
Specimen:

Bronchoalveolar lavage (BAL), nasopharygeal aspirate (NPA), or throat swab. Sputum, cerebrospinal fluid (CSF), and tissue samples may also be tested but are not ideal.  Dry swabs will not be accepted for testing; any swabs sent must be supplied in appropriate storage medium.  At least 0.5 mL of fluid sample is required for testing, and it is preferable if all samples are supplied in screw-capped tubes made of freeze-thaw and shatter-resistant plastic.

Collection Method:

Please follow standard aseptic sampling methods for collecting specimens and ensure any tissue samples/swabs are in appropriate storage medium.

Specimen Processing, Storage and Shipping:

No further processing at the sending lab is required once specimens are collected according to the above instructions.  It is preferable that all specimens be frozen after collection.  If that’s not possible, then store at refrigeration temperature (2 – 8°C).   Ensure that specimens are held at the appropriate temperature when shipped to the NML (e.g. frozen specimens shipped with dry ice, refrigerated specimens with cold packs).

Transportation of Dangerous Goods:

Shipping of specimens shall be done by a TDG certified individual in accordance with TDG regulations. For additional information regarding classification of specimens for the purposes of shipping, consult either Part 2 Appendix 3 of the TDG Regulations or section 3.6.2 of the IATA Dangerous Goods Regulations as applicable.

For additional guidance on the transport of infectious substances in other languages, please click on the link below.

http://www.who.int/ihr/capacity-strengthening/infectious-substances/en/

Patient Criteria:

Clinical features of M. pneumoniae respiratory infection frequently include such symptoms as tonsillitis, rhinitis, bronchiolitis, and atypical pneumonia, but symptoms related to other systems may occur (e.g. neurological, renal, dermatological, ophthalmological, musculoskeletal, haematological/cardiovascular, and gastrointestinal).

Accompanying Documentation:

Completed Special Bacteriology requisition form detailing all patient information and relevant clinical information. If possible, attach lab results that have already been done at local or provincial laboratories.

 

Comments:

If a test performed by the submitting lab produced a positive result for M. pneumoniae, it is also helpful to indicate this in the documentation including what type of test was performed.

Methods and Interpretation of Results:

DNA from all M. pneumoniae samples received at the NML for PCR testing is initially extracted using a commercially-available kit.  Results are based upon a PCR specific for M. pneumoniae  as well as a PCR to detect the 16S rRNA gene of the Mycoplasma genus. A sample is interpreted as positive for M. pneumoniae if the M. pneumoniae-specific PCR is positive.  If a sample is negative for M. pneumoniae PCR but positive for Mycoplasma 16S rRNA PCR, the product will be sequenced to confirm species identification at the sender’s request.

Turnaround Time:

10 calendar days. Please note that during times when large numbers of samples are received or if tests must be repeated the turnaround time may be longer.

Contact:
Phone: (204) 789-2137
Fax: (204) 784-7509
References:
  1. Sanchez-Vargas, FM and Gómez-Duarte OG. 2008. Mycoplasma pneumoniae-an emerging extra-pulmonary pathogen. Clin Microbiol Infect. Feb;14(2):105-17
  2. Lingappa JR, Lawrence W, West-Keefe S, Gautom R, and Cookson BT. 2002. Diagnosis of community-acquired pertussis infection: comparison of both culture and fluorescent-antibody assays with PCR detection using electrophoresis or dot blot hybridization. J. Clin. Microbiol. 40(8):2908-12.
  3. van Kuppeveld FJ, van der Logt JT, Angulo, AF, van Zoest MJ, Quint WG, Niesters HG, Galama JM, Melchers WJ. 1992. Genus- and species-specific identification of mycoplasmas by 16S rRNA amplification. Appl. Env. Microbiol. 58(8): 2606 – 2615.
  4. Bernet C, Garret M, de Barbeyrac B, Bebear C, and Bonnet J. 1989. Detection of Mycoplasma pneumoniae by using the polymerase chain reaction. J. Clin. Microbiol. 27(11): 2492 – 2496.
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