Order Code LABMTBPZ Mycobacterium tuberculosis Complex, Pyrazinamide Resistance by pncA DNA Sequencing, Varies
Reporting Name
Mtb PZA Resistance, pncA SequencingUseful For
Detection of genotypic resistance to pyrazinamide by Mycobacterium tuberculosis complex isolates
Performing Laboratory
Mayo Clinic Laboratories in RochesterSpecimen Type
VariesShipping Instructions
1. See Infectious Specimen Shipping Guidelines.
2. Place specimen in a large infectious container (T146) and label as an etiologic agent/infectious substance.
Necessary Information
Specimen source and suspected organism identification are required.
Specimen Required
Specimen Type: Organism
Supplies: Infectious Container, Large (T146)
Container/Tube: Middlebrook 7H10 agar slant
Specimen Volume: Isolate
Collection Instructions: Organism must be in pure culture, actively growing.
Specimen Stability Information
Specimen Type | Temperature | Time | Special Container |
---|---|---|---|
Varies | Ambient (preferred) | ||
Refrigerated |
Special Instructions
Reference Values
Pyrazinamide resistance not detected
Day(s) Performed
Varies
CPT Code Information
87153-Mtb PZA Confirmation, pncA Sequence
Method Description
Organisms identified as Mycobacterium tuberculosis complex using the Mycobacterium tuberculosis AccuProbe (GenProbe) are lysed using the PrepMan Ultra lysis buffer. Using the pncA primers described by Shenai and colleagues, an approximately 700 base pair-polymerase chain reaction (PCR) product is generated that flanks the entire pncA gene and the upstream promoter region. The PCR product is cleaned and sequenced using the Big Dye terminator v 1.1 Cycle Sequencing reagents (Applied Biosystems). Results are analyzed versus the wildtype pncA sequence using MicroSeq Microbial ID software. A custom library of non-wildtype sequences was constructed in MicroSeq. An exact match to the custom nucleotide library is required to report the result.(Shenai S, Rodrigues C, Sadani M, Sukhadia N, Mehta A: Comparison of phenotypic and genotypic methods for pyrazinamide susceptibility testing. Indian J Tuberc. 2009;56(2):82-90)
Reject Due To
Agar plate | Reject |
Reflex Tests
Test ID | Reporting Name | Available Separately | Always Performed |
---|---|---|---|
MTBVP | Mtb PZA Confirmation, pnc A Sequence | No, (Bill Only) | No |
Testing Algorithm
When this test is ordered, the reflex test may be performed and charged.
Method Name
DNA Sequencing
Forms
If not ordering electronically, complete, print, and send a Microbiology Test Request (T244) with the specimen.