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Molecular diagnostics in tuberculosis.

V C C Cheng¹, W W Yew, K Y Yuen

¹Centre of Infection and Immunology, University of Hong Kong, Hong Kong Special Administrative Region, China.

European Journal of Clinical Microbiology & Infectious Diseases : Official Publication of the European Society of Clinical Microbiology

|November 12, 2005

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Molecular diagnostics rapidly detect tuberculosis (TB) and drug resistance. A brief culture step before nucleic acid amplification tests improves sensitivity for low bacterial loads, aiding TB diagnosis.

Area of Science:

Microbiology
Molecular Diagnostics
Infectious Diseases

Background:

Molecular diagnostics offer rapid detection of Mycobacterium tuberculosis complex.
Nucleic acid amplification (NAA) tests are specific but less sensitive than culture.
Successful NAA depends on mycobacterial DNA shedding into clinical specimens.

Purpose of the Study:

To evaluate the role and limitations of NAA tests in tuberculosis diagnosis.
To address challenges with NAA sensitivity, particularly in paucibacillary cases.
To optimize NAA testing protocols for improved diagnostic accuracy.

Main Methods:

Utilized nucleic acid amplification (NAA) tests for Mycobacterium tuberculosis detection.
Compared NAA test performance with Ziehl-Neelsen smears and mycobacterial culture.

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Investigated the impact of PCR inhibitors on NAA test results.

Introduced a brief culture augmentation step prior to NAA testing.

Main Results:

NAA tests are crucial for diagnosing active tuberculosis in multibacillary disease when combined with positive smears.
NAA tests are more critical in paucibacillary tuberculosis but susceptible to false-negative results due to inhibitors.
Extra extraction steps to remove inhibitors can reduce mycobacterial DNA yield.
A brief culture augmentation step enhances mycobacterial load and dilutes inhibitors, improving NAA sensitivity.

Conclusions:

NAA tests are valuable tools in tuberculosis diagnostics, especially when sensitivity is enhanced.
Culture augmentation before NAA testing is an effective strategy to overcome inhibitor-associated false-negative results.
Optimized NAA protocols, including culture augmentation, maintain diagnostic sensitivity without significantly increasing turnaround time.