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Health Surveillance and Disease Management / Communicable Diseases / Tuberculosis

Manual for the Bacteriological Diagnosis of Tuberculosis:

Standards and Technical Guidelines—Part 2, Cultures

TB manual, cultures

Full text (in Spanish, 114 pp, PDF, 2026 Kb; chapter headings translated below for user orientation)
- Preface (text to right)
Usefulness of Cultures
- Organizing Culture Services in the Laboratory Network
- The Tuberculosis Bacillus and Other Mycobacteria Present in Human Clinical Samples
- Basis of the Procedures Utilized for Mycobacterium tuberculosis Cultures: Choosing the Culture Method in Accordance with Available Resources
- Taking, Receiving, Conserving, and Transporting Samples
- Techniques and Procedures for Sample Cultures
- Reading and Reporting Results
- Identifying Mycobacterium tuberculosis
- Quality Control (internal, external)
- Selected Bibliography
Annexes
Annex I: Minimum Biosafety Measures
Annex II: Equipment
Annex III: Plastic, Glass, and Other Materials
Annex IV: Preparing Materials, Media, and Reactives
Annex V: Models for Forms and Registries

Editorial Team
Author:
L. Barrera, National Institute of Infectious Diseases (Instituto Nacional de Enfermedades Infecciosas / INEI), "Dr. Carlos G. Malbrán" National Laboratory and Health Institutes Administration (Administración Nacional de Laboratorios e Institutos de Salud / ANLIS), Argentina
Technical Review:
M.D. Sequeira de Latini, National Institute of Respiratory Diseases (Instituto Nacional de Enfermedades Respiratorias / INER/ANLIS), Argentina
S. Balandrano, Epidemiological Diagnositic and Reference Institute (Instituto de Diagnóstico y Referencia Epidemiológicos / InDRE), Mexico
M. Velazco, Institute of Public Health (Instituto de Salud Pública / ISP), Chile
E. Montoro, "Pedro Kourí" Institute of Tropical Medicine (Instituto de Medicina Tropical "Pedro Kourí" / IPK), Cuba
Special Review by Experts:
I. Narvaiz de Kantor and A. Laszlo
Special Thanks:
M.A. Telles
M.C. Garzón Torres, National Institute of Health (Instituto Nacional de Salud / INS), Colombia
For the publications documenting the Brazilian and Colombian experiences with the Ogawa culture media and the Kudoh method
David Avendaño and Beatriz López for the photographs

Part 1, Smear Tests

PAHO Links
- Tuberculosis Page

WHO Links Stop TB
- New Technologies for TB Control: A Framework for Their Adoption, Introduction, and Implementation:
Executive Summary
- WHO TB Page

cultures

Handbook produced especially for strengthening laboratory capacity in Latin America

As was presented in the first part of this Manual, sputum-smear testing is the technique that supports tuberculosis control measures. With sputum-smear microscopy, the laboratory can start investigating a sample of patient lesions and look for the TB bacillus, detect it, evaluate the evolution of infectious cases, predict and endorse a cure for patients who successfully complete their treatment regimen, and identify those who fail in their treatment.

  • Cultures produce results later but are more sensitive than sputum-smear microscopy.
  • A culture can show a minimum of 10–100 acid-fast bacilli (AFB) present in a sample, if carried out properly.
  • Cultures allow for detecting cases before they become infectious.

Cultures complement smear tests, since they can show viable bacilli present in limited quantity in a lesion sample, characterizing them so as to ascertain that it is indeed the TB bacillus and see if it is sensitive or resistant to TB drugs. Hence, the role of cultures is more important in scenarios with a median or low incidence of tuberculosis, with high levels of TB/HIV co-infection, and with a multiresistant medium or a high tuberculosis burden. Molecular techniques have been developed to achieve the same results of cultures with greater accuracy. These methods have still not managed to replace culture-dependent detection, identification, and sensitivity testing. A culture can be applied in laboratories with medium-level resources and maintains its position as main reference method by virtue of its precision. On the other hand, no immunological method has managed to equate the specificity of the culture in detecting and confirming the diagnosis of the disease caused by the TB bacillus; subsequently, on the basis of evidence to date, it is not reasonable to introduce such innovations into network tuberculosis diagnostic laboratories without ensuring beforehand coverage and quality with sputum-smear testing and cultures.

Considering the prevalent epidemiological situation Latin America, the experience developed in the Region and the existing resources, it is appropriate to promote the value of cultures as the tool for tuberculosis control programs. A culture can improve program efficiency evaluations in treatment administration, optimize management of multidrug-resistant tuberculosis and that associated with HIV, and help make progress in controlling the disease where objectives established for infectious cases have been reached.

Selective Use of Cultures

Upon diagnosis: Cultivate all samples of symptomatic patients with clinical signs and/or x-ray or other images compatible with tuberculosis and some of the following characteristics:

  • negative sputum-smear test from three respiratory samples
  • extrapulmonary localization of the disease
  • children
  • immuno-supressed patients, particularly HIV-positive
  • positive sputum-smear test in gastric lavage, bronchial wash, or swabs
  • background of tuberculosis treatment, especially if abandonment or treatment failure was recorded
  • exposure to drug-resistant bacillus infection (contacts with drug-resistant tuberculosis cases, inpatients or health workers from institutions or prisons where there are recorded cases of multidrug-resistant tuberculosis)

During treatment control: Cultivate samples of

  • chronic tuberculosis cases or those with positive sputum-smear microscopy in controls from the second month of chemotherapy or later
  • cases diagnosed with a negative sputum-smear microscopy and who convert to a positive sputum-smear test during treatment

For surveillance of resistance to anti-tubercular drugs :

  • Cultivate the samples of cases under study or surveillance as provided for in work protocol

The situation analysis also leads to promoting quality assurance of cultures and ensuring access to this technique on the part of patients who can benefit from it.

With this in mind, we have updated the standards for the identifying Mycobacterium tuberculosis through cultures. This second part of the Manual on Tuberculosis Bacteriology is devoted to laboratories that are assumed to already have ensured competition, quality, and biosafety in performing sputum-smear tests. Hence, the procedures presented here complement those described for carrying out sputum-smear microscopy in the first part of the Manual.

These standards have taken into account those preceding them in Latin America (CEPANZOS Technical Notes No. 26, 27, 28 and 29/1988), those prepared by WHO (WHO/TB/98,258), and the experiences of the laboratories of the Region. They propose improving the technique implemented by the previous standard and offer a menu of methods for different scenarios, selected from those that have proved to contribute to TB diagnosis in Latin America, according to the evidence published. In addition, they provide details on procedures to increase the quality and biosafety in the workplace.

Stop TB / Alto a la TB