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Related Experiment Video

Updated: Jun 21, 2026

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

Published on: April 25, 2014

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

E Flores¹, J C Rodríguez, E Garcia-Pachón

¹Section of Microbiology, Hospital General Universitario, Universidad Miguel Hernández, Elche, Spain.

APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica

|August 12, 2009

Summary

A new real-time PCR test for tuberculosis diagnosis now includes an internal amplification control (IAC). This validated assay enhances M. tuberculosis detection and aids in standardizing diagnostic methods.

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Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

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Last Updated: Jun 21, 2026

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

Published on: April 25, 2014

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Development of a Quantitative Recombinase Polymerase Amplification Assay with an Internal Positive Control

Published on: March 30, 2015

Area of Science:

Microbiology
Molecular Diagnostics
Infectious Disease

Background:

Real-time PCR is crucial for tuberculosis diagnosis.
Current tests often lack an internal amplification control (IAC), limiting clinical utility.
A validated real-time PCR with IAC is needed for improved tuberculosis detection.

Purpose of the Study:

To design and validate a novel, user-friendly real-time PCR assay for tuberculosis diagnosis.
To incorporate an internal amplification control (IAC) for enhanced reliability.
To assess the performance of the assay in clinical samples.

Main Methods:

Developed a real-time PCR targeting the IS6110 of Mycobacterium tuberculosis.
Designed an IAC using a fragment of Chlamydia trachomatis.
Validated the assay using clinical samples including urine, sputum, and isolates of non-tuberculous mycobacteria.

Main Results:

The assay demonstrated very low interassay (0.45-1.65%) and intraassay (0.18-1.80%) variation.
Achieved 100% specificity in detecting M. tuberculosis.
The detection limit was approximately 15 bacteria without IAC and 32 bacteria with IAC.

Conclusions:

The developed real-time PCR with IAC assay improves M. tuberculosis detection accuracy.
The assay contributes to the standardization of tuberculosis diagnostic techniques.
This method offers a reliable tool for clinical application in tuberculosis diagnosis.