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Order Code LABMTBPZ Mycobacterium tuberculosis Complex, Pyrazinamide Resistance by pncA DNA Sequencing, Varies

Infectious

Reporting Name

Mtb PZA Resistance, pncA Sequencing

Useful For

Detection of genotypic resistance to pyrazinamide by Mycobacterium tuberculosis complex isolates

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Varies


Shipping 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 

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.