Introduction to the course
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Agenda
Include:
TECHNICAL ISSUES
PRACTICAL ISSUES
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TB situation
• TB is the typical disease of poverty. Of the 22
countries that have 80% of the world's TB
burden, 17 are classified as low-income
countries.
• The lack of human resources available for TB
control at all levels is a global problem.
New donors and opportunities for
international collaboration have become
available as recently highlighted by the
launch of the Global Laboratory Initiative
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General objectives
• Train laboratory personnel:
‒ in performing M. tuberculosis culture and DST;
‒ in working safely in a laboratory performing
cultures and DST;
‒ in complying with quality assurance requirements.
• Raise awareness of the importance of
laboratory performance in TB control.
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Target audience
• Laboratory personnel who will perform
specimen processing, cultures of tubercle bacilli
and drug susceptibility testing of isolated strains
in the laboratory component of the national TB
control programme (or other sector involved in
TB diagnosis).
• The course is intended for laboratory personnel
in low- and middle-income countries working in
laboratories at the central and intermediate
levels.
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Requirements for trainees
• Proficient in TB smear microscopy and handling of
specimens (recognize good-quality samples, use registers,
aware of consequences of laboratory errors for patient
management).
• Trained in good laboratory practice (familiarity with
laboratory environment and differentiation between
clean/dirty working areas; use and maintenance of standard
laboratory equipment such as balance, incubator, freezer,
refrigerator) and used to following written protocols.
• Familiar with quality control (QC) and external quality
assurance (EQA) for smear microscopy.
• Aware of the national TB control programme structure and
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laboratory network.
Course methodology
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Lessons
Interactive
Exercise/experience/problem-solving
Practical work in the laboratory
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Materials provided
• Participant’s manual, in modules (folder)
• Agenda (folder)
• PowerPoint presentations and exercise
slides (handouts and CD)
• Technical materials (CD)
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Learning objectives ‒ Module 1
Overview: Tuberculosis, the global emergency
At the end of this module, participants will be able to:
 comment on the worldwide and local TB epidemic;
 explain TB/HIV and MDR- XDR-TB;
 describe the forms and transmission of TB;
 describe the new Stop TB Strategy;
 explain the roles of the NTP and the laboratory network;
 explain the importance of AFB microscopy, culture and
DST in TB control.
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Learning objectives ‒ Module 2
Safety precautions for tuberculosis culture
and DST
At the end of this module, participants will be able to:
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explain airborne transmission of TB;
adhere to safe practice;
explain the biosafety levels of TB laboratories;
work safely in biological safety cabinets;
use adequate protective personal equipment;
use appropriate disinfectants;
cope with accidents in the TB laboratory;
handle chemicals safely in the laboratory .
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Learning objectives ‒ Module 3
Use, calibration and maintenance of
laboratory equipment
At the end of this module participants will be able to:
 describe briefly the operating characteristics of biological
safety cabinets, centrifuges, autoclaves, inspissators, pH
meters, incubators;
 correctly use biological safety cabinets, centrifuges,
autoclaves, inspissators, pH meters, incubators;
 describe the main maintenance operations for biological
safety cabinets, centrifuges, autoclaves, inspissators, pH
meters, incubators.
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Learning objectives ‒ Module 4
Preparation of solid media for culture and DST
At the end of this module participants will be able to:
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recognize different media for mycobacteria culture and
explain their advantages/disadvantages;
prepare all the reagents, including drug solutions, for
preparation of media for culture and DST;
prepare and dispense the culture medium;
check the quality of tubes at the end of the process
and store properly;
perform the sterility check.
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Learning objectives ‒ Module 5
Collection, transportation and receipt of
specimens
At the end of this module participants will be able to:
 explain the proper collection of specimens;
 explain how to select suitable containers for the
collection of different specimens;
 explain the proper labelling of specimens to be submitted
to the TB laboratory;
 explain the proper transportation of specimens;
 list the features of a good specimen;
 fill in the laboratory register.
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Learning objectives ‒ Module 6
Processing the specimens and inoculation of
solid and liquid media
At the end of this module participants will be able to:
 work properly within a BSC;
 Process specimens from sterile and non sterile sites,
according to protocols, for TB culture;
 homogenize and decontaminate respiratory specimens
by:
– Petroff modified method,
– NALC-NaOH method;
– Kudoh method;
 inoculate cultures (solid and/or liquid media);
 incubate inoculated media under proper conditions.
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Learning objectives ‒ Module 7
Reading cultures
At the end of this module participants will be able to:
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examine cultures at appropriate times;
identify presumptive M. tuberculosis colonies;
recognize contaminations;
report the suspect positive cultures.
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Learning objectives ‒ Module 8
Identification of M. tuberculosis
At the end of this module participants will be able to:
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evaluate growth characteristics for species identification;
perform and interpret the PNB growth inhibition test;
perform and interpret the niacin test
perform and interpret the nitrate reduction test
 perform and interpret the catalase test:
– heat-labile;
 Understand the principle of immunochromatographic tests.
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Learning objectives ‒ Module 9
Recording and reporting culture results
At the end of this module participants will be
able to:
 record culture results in the laboratory
register;
 record and report accordingly
(semiquantitative) on WHO forms.
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Learning objectives ‒ Module 10
Performing drug susceptibility testing
At the end of this module participants will be able to:
 understand the objectives and rationale of combined anti-TB therapy;
 understand the basic definitions of drug resistance in TB;
 understand how drug resistance develops;
 understand the rationale for using a critical concentration of drug in
laboratory assays;
 understand the rationale for determining the proportion of resistant
bacteria that is clinically significant;
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explain the different methods for performing DST;
perform and interpret the proportion method;
record and report DST results;
store strains.
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Learning objectives ‒ Module 11
Compliance with established quality assurance
requirements
At the end of this module participants will be able to:
 explain the three main components of the QA system
and their importance;
 adhere to the QC and EQA procedures.
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Learning objectives ‒ Module 12
Managing supplies for a tuberculosis laboratory
performing culture
At the end of this module, participants will be able to:
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list supplies required for culture;
explain ordering of supplies;
maintain proper records;
explain use of the stock book;
calculate supplies required;
inspect and verify supplies required;
explain storage of supplies.
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Course evaluation
• Self-evaluation by suggested key
questions at the end of each module
• Evaluation by participants (questionnaire)
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