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Laboratory Diagnosis and Susceptibility Testing for

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  • Author: Gary W. Procop1
  • Editor: David Schlossberg2
  • VIEW AFFILIATIONS HIDE AFFILIATIONS
    Affiliations: 1: Department of Laboratory Medicine, Cleveland Clinic, Cleveland, OH 44195; 2: Philadelphia Health Department, Philadelphia, PA
  • Source: microbiolspec December 2016 vol. 4 no. 6 doi:10.1128/microbiolspec.TNMI7-0022-2016
  • Received 17 October 2016 Accepted 20 October 2016 Published 09 December 2016
  • Gary W. Procop, procopg@ccf.org
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  • Abstract:

    The laboratory, which utilizes some of the most sophisticated and rapidly changing technologies, plays a critical role in the diagnosis of tuberculosis. Some of these tools are being employed in resource-challenged countries for the rapid detection and characterization of . Foremost, the laboratory defines appropriate specimen criteria for optimal test performance. The direct detection of mycobacteria in the clinical specimen, predominantly done by acid-fast staining, may eventually be replaced by rapid-cycle PCR. The widespread use of the Xpert MTB/RIF (Cepheid) assay, which detects both and key genetic determinants of rifampin resistance, is important for the early detection of multidrug-resistant strains. Culture, using both broth and solid media, remains the standard for establishing the laboratory-based diagnosis of tuberculosis. Cultured isolates are identified far less commonly by traditional biochemical profiling and more commonly by molecular methods, such as DNA probes and broad-range PCR with DNA sequencing. Non-nucleic acid-based methods of identification, such as high-performance liquid chromatography and, more recently, matrix-assisted laser desorption/ionization–time of flight mass spectrometry, may also be used for identification. Cultured isolates of should be submitted for susceptibility testing according to standard guidelines. The use of broth-based susceptibility testing is recommended to significantly decrease the time to result. Cultured isolates may also be submitted for strain typing for epidemiologic purposes. The use of massive parallel sequencing, also known as next-generation sequencing, promises to continue to this molecular revolution in mycobacteriology, as whole-genome sequencing provides identification, susceptibility, and typing information simultaneously.

  • Citation: Procop G. 2016. Laboratory Diagnosis and Susceptibility Testing for . Microbiol Spectrum 4(6):TNMI7-0022-2016. doi:10.1128/microbiolspec.TNMI7-0022-2016.

Key Concept Ranking

Restriction Fragment Length Polymorphism
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References

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2016-12-09
2017-08-17

Abstract:

The laboratory, which utilizes some of the most sophisticated and rapidly changing technologies, plays a critical role in the diagnosis of tuberculosis. Some of these tools are being employed in resource-challenged countries for the rapid detection and characterization of . Foremost, the laboratory defines appropriate specimen criteria for optimal test performance. The direct detection of mycobacteria in the clinical specimen, predominantly done by acid-fast staining, may eventually be replaced by rapid-cycle PCR. The widespread use of the Xpert MTB/RIF (Cepheid) assay, which detects both and key genetic determinants of rifampin resistance, is important for the early detection of multidrug-resistant strains. Culture, using both broth and solid media, remains the standard for establishing the laboratory-based diagnosis of tuberculosis. Cultured isolates are identified far less commonly by traditional biochemical profiling and more commonly by molecular methods, such as DNA probes and broad-range PCR with DNA sequencing. Non-nucleic acid-based methods of identification, such as high-performance liquid chromatography and, more recently, matrix-assisted laser desorption/ionization–time of flight mass spectrometry, may also be used for identification. Cultured isolates of should be submitted for susceptibility testing according to standard guidelines. The use of broth-based susceptibility testing is recommended to significantly decrease the time to result. Cultured isolates may also be submitted for strain typing for epidemiologic purposes. The use of massive parallel sequencing, also known as next-generation sequencing, promises to continue to this molecular revolution in mycobacteriology, as whole-genome sequencing provides identification, susceptibility, and typing information simultaneously.

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Figures

Image of FIGURE 1
FIGURE 1

The colonies of two different species are demonstrated on L-J agar slants. The isolate on the left is “rough and buff,” with colonies that have a cauliflower-like appearance. The isolate on the right, for comparison, is , a photochromogen that demonstrates pigment development after exposure to light.

Source: microbiolspec December 2016 vol. 4 no. 6 doi:10.1128/microbiolspec.TNMI7-0022-2016
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Image of FIGURE 2
FIGURE 2

The MGIT 960 tube on the right contains growing mycobacteria and is fluorescent when exposed to UV light. In contrast, the tube on the left contains no mycobacteria.

Source: microbiolspec December 2016 vol. 4 no. 6 doi:10.1128/microbiolspec.TNMI7-0022-2016
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Image of FIGURE 3
FIGURE 3

The aggregation of acid-fast bacilli into the suprastructure demonstrated here is termed cording and is highly suggestive of complex. The culture from which this was derived contained complex. The image is of a Ziehl-Neelsen stain at a magnification of ×500.

Source: microbiolspec December 2016 vol. 4 no. 6 doi:10.1128/microbiolspec.TNMI7-0022-2016
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Image of FIGURE 4
FIGURE 4

These postamplification melting curves, derived using fluorescence resonance energy transfer probes following broad-range mycobacterial PCR, demonstrate that complex (MTB) can be differentiated from the nontuberculous mycobacteria (MK), (MA), and (MI).

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Image of FIGURE 5
FIGURE 5

This pyrogram, derived from pyrosequencing of the hypervariable A region of the 16S rRNA gene following a broad-range mycobacterial PCR, may be used for mycobacterial identification.

Source: microbiolspec December 2016 vol. 4 no. 6 doi:10.1128/microbiolspec.TNMI7-0022-2016
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