Conventional culture methods may take up to six weeks to detect growth of M.tuberculosis, whereas NAAT can be performed directly on sputum and other respiratory specimens and results can be provided to the clinician within 24-48 hours of specimen receipt. In January 2009 CDC published Updated Guidelines for the Use of Nucleic Acid Amplification Tests in the Diagnosis of Tuberculosis.
The Centers for Disease Control and Prevention (CDC) recommends in its 2009 guidance that NAAT be performed on at least one respiratory specimen from each patient with signs and symptoms of pulmonary TB for whom a diagnosis of TB is being considered but has not yet been established, and for whom the test result would alter case management or TB control activities. TB NAAT should only be ordered on patients when the clinician’s index of suspicion for pulmonary TB is moderate to high (more).
The test should not be ordered when the clinician has a low index of suspicion for pulmonary TB in the patient, as the positive predictive value of the test falls to <50% if ordered under this circumstance (1)
The Rhode Island State Health Laboratory accepts only the following respiratory specimens for TB NAAT: sputum, bronchoalveolar lavage (BAL), bronchial washing, bronchial lavage, lung washing, thoracentesis fluid, and pleural fluid. Our NAAT PCR method has been verified on these specimens in an extensive in-house study.
To date there is limited information regarding TB NAAT performance on non-respiratory specimens or specimens from patients under treatment. (1)
The State Health Laboratory performs TB NAAT by real time PCR. The test was developed “in-house” and is a modification of the PCR method developed, validated and currently used by the New York State Wadsworth Center. The test is not FDA approved, but has been validated and verified in accordance with applicable Clinical Laboratory Improvement Amendments (CLIA) regulations.
Yes, AFB smear and culture should always be ordered on specimens submitted for NAAT. The result of the NAAT is always interpreted in conjunction with the AFB smear result and AFB culture remains the “gold standard” for the laboratory confirmation of TB and is required in order to conduct drug susceptibility testing and genotyping. When a specimen is sent to our laboratory for a TB NAAT, an AFB smear and a culture will also be ordered and performed.
The Rhode Island State Health Laboratory will require an additional unprocessed specimen be submitted to our laboratory for NAAT. This same specimen will also have an AFB smear and culture done at our laboratory. The method verification that took place in our laboratory was done on specimens that had been through the specimen decontamination process used in our laboratory, so it has not been verified for use on specimens that have been decontaminated by other methods.
If a hospital laboratory wishes to send the remaining sediment from a specimen that they have all ready decontaminated, along with the additional unprocessed specimen, we would encourage them to do so. This will allow our laboratory to do additional verification studies to see if other decontamination methods are compatible with our NAAT PCR method.
Complete both a Rhode Island State Health Laboratory Test Requisition Form and a TB NAAT Request Form. Both forms are available in all hospital microbiology and send out laboratories and can also be requested by contacting the Rhode Island State Health Laboratory Mycobacteriology Lab at 222-5587/222-5586.
The Mycobacteriology section of the Special Pathogens Lab processes approximately 2,000 specimens a year for AFB smear and culture. If we determine that a specimen (acceptable for NAAT) is AFB smear positive, we will automatically order a TB NAAT provided:
Our goal is to have NAAT results available within two working days of specimen receipt. Specimens should be sent to the State Health Laboratory as soon as possible after collection.
All positive TB NAAT results will be considered a critical test value and will be immediately phoned to the ordering provider, the laboratory that submitted the specimen, the Health Department Tuberculosis Control Program, and the RISE TB Clinic. NAAT results will be routinely reported through our laboratory information system to submitters in the same manner they receive other reports from our laboratory.
No, a single negative NAAT should not be used as a definitive result to exclude TB (refer to NAAT Interpretation Guidelines) especially in a patient where the clinical suspicion of TB is moderate to high. If the clinician is not experienced with the interpretation of NAAT or the diagnosis of TB, consultation with the expert TB clinicians at the RISE TB Clinic is highly recommended (401) 793-2427/2433. (1)
A negative NAAT and two independent AFB smear-negative sputum samples (taken at least 8 hours apart, with at least one being an early morning specimen) may be used to determine whether patients may be removed from respiratory isolation, provided that (a) clinical suspicion for infectious TB is low and (b) if the patient has been started on multidrug treatment, there is clinical improvement. (1)