Molecular Detection of Mycobacterium leprae by real-time PCR
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Reference Details
Real-time PCR based detection of Mycobacterium species including M. leprae in clinical samples using various molecular targets.
- Leprosy
Please consult with the NRCM Diagnostic Head and refer below to determine if your sample is appropriate for this type of testing. Please note the volume and quality of sample for testing:
- For CSF, or other fluid submissions: Optimally 5 mL, or a minimum of 1 mL sample.
- For tissues: Fresh sterile samples, including fresh biopsies, are preferred. Sample should be AFB positive. Frozen samples may be accepted as long as they have been handled in an aseptic environment only.
- For DNA: A minimum of 100 ng of DNA is required. The DNA should be suspended in a minimum volume of 25 ul molecular grade water/buffer along with a quantification of the DNA in the samples.
Clinical samples that are positive for acid fast bacilli such as fresh tissue, CSF or sediment from decontaminated samples. Formalin fixed tissues and/or paraffin-embedded samples are not recommended for testing due to DNA degradation.
Ship clinical specimens such as sputum or fresh tissue on ice for overnight delivery, and prior to Wednesday each week to ensure receipt by Friday. Previously frozen material can be shipped on dry ice using appropriate Transportation of Dangerous Goods shipping procedures. Please consult the NRCM if further information is required.
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.
Please consult with the NRCM if you have any questions prior to submitting a sample for M. leprae detection. Please note patient travel history.
Requisitions must be emailed or faxed prior to shipping isolates. Samples should be shipped attention: Melissa Rabb, Head, Reference and Diagnostic Services, NRCM/NML, ph: 204-789-6081/204-789-6038. The requisition for the NRCM should be completed and signed off by the supervisor/designate and include the submitting laboratory identifier (specimen ref #), patient gender, year of birth, clinical history, travel history, source of specimen, submitter information on microscopy if available, and any other information relevant to the submitted sample. Please request M. leprae detection in the Identification and Detection section of the NRCM requisition.
Samples may be rejected if a low volume of sample is submitted or appropriate documentation is incomplete or missing.
Real-time PCR will be employed for the detection of the unique RLEP repeat sequence and the ITS region for M. leprae testing.
10 calendar days.
- Truman RW, Andrews PK, Robbins NY, Adams LB, Krahenbuhl JL, Gillis TP. Enumeration of Mycobacterium leprae using real-time PCR. PLoS Negl Trop Dis. 2008;2: e328. doi:10.1371/journal.pntd.0000328.