Real-time PCR with internal amplification control for detecting tuberculosis: method design and validation

$48.00 plus tax (Refund Policy)

Download / Buy Article:


Flores E, Rodríguez JC, Garcia-Pachón E, Soto JL, Ruiz M, Escribano I, Royo G. Real-time PCR with internal amplification control for detecting tuberculosis: method design and validation. APMIS 2009; 117: 592–7.

Real-time PCR has been a major development in the diagnosis of tuberculosis. However, most tests do not include an internal amplification control (IAC), which therefore limits it clinical application. In this study a new, easy to perform real-time PCR test with IAC was designed and validated in clinical samples. The primers amplified a 163-bp fragment of IS6110 of Mycobacterium tuberculosis and the IAC was designed with a fragment of a different microorganism (Chlamydia trachomatis). The interassay and intraassay variation of this test were very low (0.45–1.65% and 0.18–1.80%, respectively). The detection accuracy was validated in 50 samples (25 urine, 25 sputum) with different concentrations of M. tuberculosis, 18 clinical isolates of non-tuberculous mycobacteria and 148 samples with clinical suspicion of pulmonary tuberculosis. The specificity was 100%. The detection limit of this PCR test without IAC was approximately 15 bacteria and with IAC approximately 32 bacteria. This real-time PCR with IAC assay can improve the detection of M. tuberculosis and contribute to standardization of this diagnostic technique.

Keywords: Mycobacterium tuberculosis; diagnostic; internal control; real-time PCR

Document Type: Research Article


Affiliations: 1: Section of Microbiology 2: Section of Pneumology 3: Research Unit, Hospital General Universitario, Universidad Miguel Hernández, Elche, Spain

Publication date: August 1, 2009

Related content



Share Content

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
Cookie Policy
ingentaconnect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more