Diagnostic Mycobacteriology Laboratory Practices

Shinnick, Thomas M.; Good, Robert C.

doi:10.1093/clinids/21.2.291

Cited by 87 publications

(52 citation statements)

References 41 publications

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“…HPLC analysis of mycolic acids for species identification offered a reprieve from the traditional time-consuming identification process (55,84 be analyzed in hours by HPLC, compared to weeks or longer for routine methods. However, the suitability of the method should be evaluated prior to incorporation into the laboratory.…”

Section: Laboratorymentioning

confidence: 99%

Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification ofMycobacteriumSpecies

2001

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Mycobacterium tuberculosis is the etiologic agent of tuberculosis and can be accurately detected by laboratories using commercial genetic tests. Nontuberculosis mycobacteria (NTM) causing other mycobacterioses can be difficult to identify. The identification processes are confounded by an increasing diversity of newly characterized NTM species. The ubiquitous nature of NTM, combined with their potential to be opportunistic pathogens in immunocompromised as well as nonimmunodeficient patients, further complicates the problem of their identification. Since clinical case management varies depending on the etiologic agent, laboratories must identify the species in a timely manner. However, only a few identification methods can detect the species diversity within the Mycobacterium genus. Over the last decade, high-performance liquid chromatography analysis of the mycolic acids has become an accepted method for identification of mycobacteria. In this review, we assess its development and usefulness as an identification technique for Mycobacterium species

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Section: Laboratorymentioning

confidence: 99%

Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification ofMycobacteriumSpecies

2001

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“…Commercial nonradiolabeled probes are available for identifying isolates of M. tuberculosis, M. gordonae, M. kansasii, and M. avium-intracellulare complex. When performed correctly, they can be highly sensitive and specific but do require approximately 10 5 to 10 6 CFU to determine conclusive results (10,24). With the use of isolates from solid media, HPLC examines the mycolic acid fingerprint patterns that differ among most species or complexes of mycobacteria.…”

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confidence: 99%

Identification of Mycobacterium spp. by Using a Commercial 16S Ribosomal DNA Sequencing Kit and Additional Sequencing Libraries

et al. 2002

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Current methods for identification of Mycobacterium spp. rely upon time-consuming phenotypic tests, mycolic acid analysis, and narrow-spectrum nucleic acid probes. Newer approaches include PCR and sequencing technologies. We evaluated the MicroSeq 500 16S ribosomal DNA (rDNA) bacterial sequencing kit (Applied Biosystems, Foster City, Calif.) for its ability to identify Mycobacterium isolates. The kit is based on PCR and sequencing of the first 500 bp of the bacterial rRNA gene. One hundred nineteen mycobacterial isolates (94 clinical isolates and 25 reference strains) were identified using traditional phenotypic methods and the MicroSeq system in conjunction with separate databases. The sequencing system gave 87% (104 of 119) concordant results when compared with traditional phenotypic methods. An independent laboratory using a separate database analyzed the sequences of the 15 discordant samples and confirmed the results. The use of 16S rDNA sequencing technology for identification of Mycobacterium spp. provides more rapid and more accurate characterization than do phenotypic methods. The MicroSeq 500 system simplifies the sequencing process but, in its present form, requires use of additional databases such as the Ribosomal Differentiation of Medical Microorganisms (RIDOM) to precisely identify subtypes of type strains and species not currently in the MicroSeq library.

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“…Patients unable to produce sputum spontaneously should undergo sputum induction, which requires inhalation of an aerosol of sterile hypertonic saline (3%-15%) in negative-pressure isolation rooms. 70,71 Bronchoscopy with bronchoalveolar lavage may be necessary in patients unable to produce adequate expectorated or induced sputum samples. Nucleic acid amplification (NAA) testing should be performed on at least 1 respiratory specimen.…”

Section: Diagnosismentioning

confidence: 99%

Current Concepts in the Management of Tuberculosis

Sia

Wieland

2011

Mayo Clinic Proceedings

160

108

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E ffective medical therapy for tuberculosis (TB) has existed for more than half a century, yet TB remains among the most pressing public health issues of our day. Tuberculosis is, in part, a disease of poverty. 1 The fact that it remains the eighth leading cause of death in the world speaks to the challenges facing practitioners and public health officials as they try to control a disease that is so entwined in the cultural and economic fabric of society. Challenges to effective solutions include lack of access to diagnosis and treatment, the frequent coexistence of epidemics of TB and human immunodeficiency virus (HIV), and the Tuberculosis (TB) poses a serious threat to public health throughout the world but disproportionately afflicts low-income nations. Persons in close contact with a patient with active pulmonary TB and those from endemic regions of the world are at highest risk of primary infection, whereas patients with compromised immune systems are at highest risk of reactivation of latent TB infection (LTBI). Tuberculosis can affect any organ system. Clinical manifestations vary accordingly but often include fever, night sweats, and weight loss. Positive results on either a tuberculin skin test or an interferon-γ release assay in the absence of active TB establish a diagnosis of LTBI. A combination of epidemiological, clinical, radiographic, microbiological, and histopathologic features is used to establish the diagnosis of active TB. Patients with suspected active pulmonary TB should submit 3 sputum specimens for acidfast bacilli smears and culture, with nucleic acid amplification testing performed on at least 1 specimen. For patients with LTBI, treatment with isoniazid for 9 months is preferred. Patients with active TB should be treated with multiple agents to achieve bacterial clearance, to reduce the risk of transmission, and to prevent the emergence of drug resistance. Directly observed therapy is recommended for the treatment of active TB. Health care professionals should collaborate, when possible, with local and state public health departments to care for patients with TB. Patients with drug-resistant TB or coinfection with human immunodeficiency virus should be treated in collaboration with TB specialists. Public health measures to prevent the spread of TB include appropriate respiratory isolation of patients with active pulmonary TB, contact investigation, and reduction of the LTBI burden. Proc. 2011;86(4):348-361 AFB = acid-fast bacilli; BCG = bacille Calmette-Guérin; CFP-10 = culture filtrate protein 10; DOT = directly observed therapy; EMB = ethambutol; ESAT-6 = early-secreted antigenic target 6; HIV = human immuno deficiency virus; IFN-γ = interferon-γ; IGRA = IFN-γ release assay; INH = isoniazid; LTBI = latent TBI; MDR-TB = multidrug-resistant TB; NAA = nucleic acid amplification; PZA = pyrazinamide; RIF = rifampin; TB = tuberculosis; TBI = TB infection; TST = tuberculin skin test; XDR-TB = extensively drug-resistant TB Mayo Clin

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Diagnostic Mycobacteriology Laboratory Practices

Cited by 87 publications

References 41 publications

Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification ofMycobacteriumSpecies

Mycolic Acid Analysis by High-Performance Liquid Chromatography for Identification ofMycobacteriumSpecies

Identification of Mycobacterium spp. by Using a Commercial 16S Ribosomal DNA Sequencing Kit and Additional Sequencing Libraries

Current Concepts in the Management of Tuberculosis

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