BASIC ORGANIZATION
DIAGNOSTIC MICROBIOLOGY LABORATORY
Dr.T.V.Rao MD
DR.T.V.RAO MD
12-10-2012
Challenges in Diagnostic Laboratories
The work environment has changed with the development of new technology. Laboratories have always seen the need for change and development, there has been increased pressure to improve performance, tighten margins, improve quality and reduce costs
DR.T.V.RAO MD 12-10-2012
Medical Laboratory Managers
Each laboratory must have a strategic plan that describes its long-term goals, such as a move toward more automation or molecular diagnostic techniques. Each employees role should be clearly defined, and written job descriptions should be provided so personnel know what they are expected to do. Therefore, it is a not an easy task for a manger to strike a balance among the clinical laboratory regulations, fiscal responsibility, and employee competence and morale to maintain the overall quality of patient care.
DR.T.V.RAO MD 12-10-2012
THE LABORATORIES SHOULD FACTION FROM SENSE TO COMMON SENSE
It is appropriate to remember that the two most important components of management are
Common sense Open communication with laboratory staff
DR.T.V.RAO MD
12-10-2012
MICROBIOLOGY LABORATORY
Clinical Microbiology comprises essentially seven sections. Aerobic and anaerobic bacteriology Mycology
Mycobacteriology (also called Acid-fast Bacteriology, AFB)
Parasitology
Virology Serology Molecular diagnostics (PCR & DNA probe technology)
:
DR.T.V.RAO MD 12-10-2012
PLAN FOR THE DIVISION OF MICROBIOLOGY
Sample Receiving & Processing Section Samples brought to the clinical microbiology are at first received by this section. Here sample are received and the samples are processed according to the nature of the sample.
Urinalysis Section In this section detailed report of urine samples including physical, chemical, microscopic examination is be prepared.
DR.T.V.RAO MD
12-10-2012
DIVISION OF PARASITOLOGY
Parasitology Section Parasitology section deals with intestinal parasites. Samples of faeces are examined here for the presence of any intestinal parasite. Slides are prepared here inside a safety cabinet.
DR.T.V.RAO MD 12-10-2012
BACTERIOLOGY CULTURES MAIN PART OF LABORATORY SERVICES
Nose, Throat, Sputum and Urogenital Cultures and Sensitivity Section Here cultures of respiratory tract and genital tract infections are prepared.
Blood cultures, Wounds Culture and Sensitivity Section Culture of wound swab, pus, aspirates, body fluids including CSF are the responsibility of this section.
DR.T.V.RAO MD
12-10-2012
BACTERIOLOGICAL CULTURES IS THE MAJOR GRATIFYING WORK
Urine Culture & Sensitivity Section Different types of urine culture performed here including mid stream urine and catheter samples of urine. Each sample is processed and evaluated accordingly.
Blood Culture and Sensitivity Section In this section culturing of blood samples is carried out. Nowadays, this section is equipped by machines such as Bactec 9240, flourometric instruments. Each instrument is capable of running 240 samples at a time.
DR.T.V.RAO MD
12-10-2012
SEROLOGY SECTION
Serology Section In serology section immunological and serological tests are performed by different techniques like Latex agglutination, haemagglutination and antibody absorption.
DR.T.V.RAO MD
12-10-2012
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MYCOBACTERIOLOGY
Mycobacteriology Culture & Sensitivity Section In this section all TB smear, culture and sensitivity performed in two Biosafety II and III cabinets to avoid risk of infection
DR.T.V.RAO MD
12-10-2012
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MYCOLOGY LABORATORY SERVICES AN EMERGING NEED
Mycology Culture Section Requests for fungus smear and culture processed here in a bio safety II cabinet to avoid infection from fungal spore.
Regardless of the organisation of a Microbiology Laboratory, the main aim is providing the client (the physician) with accurate and reliable results to assist the process of clinical treatment.
DR.T.V.RAO MD 12-10-2012
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QUALITY CONTROL IN MICROBIOLOGY LABORATORY
Quality Control Section In this section quality control of water, food products and environment is performed with the help of different media and colony counters.
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DR.T.V.RAO MD
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MANAGING THE SAMPLE: TECHNICAL
Macroscopic evaluation Split sample for different laboratory disciplines Two possible approaches: perform only those tests requested by sender perform diagnostics for syndromes/clinical description (laboratory initiative) Storage of samples
refrigerator or freezer
DR.T.V.RAO MD
12-10-2012
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DIRECT METHODS
1. Macroscopic evaluation
2. Direct microscopy
3. Electron microscopy 4. Staining 5. Rapid tests 6. Molecular methods
No propagation required
7. Propagate the agent
DR.T.V.RAO MD
12-10-2012
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MACROSCOPIC EVALUATION AN EXAMPLE
Consistency rice water stools for Cholera
Blood
Visible parasites
Helminths
segments
DR.T.V.RAO MD 12-10-2012
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DIRECT MICROSCOPY
Wet mount technique hanging drop Dark background microscope fragile organisms (e.g. spirochetes) Viability maintained mobility may be observed
Observations
white blood cells (denotes invasion)
red blood cells parasites protozoa Helminths eggs moving bacteria
DR.T.V.RAO MD
12-10-2012
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STAINING
A specific staining
Gram staining
Specific staining with chemicals Ziehl Neelsen staining (Mycobacteria)
Modified Ziehl Neelsen staining (Cryptosporidium)
Specific staining with labelled antibodies Immunofluorescence - used when gram stain cannot help in diagnosis (e.g. Legionella too small to be visible in a Gram stain)
DR.T.V.RAO MD 12-10-2012
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RAPID TESTS
Goals
bacterial, viral or parasite antigen (surface antigen, soluble antigen)
toxin in biological fluids (e.g. cerebrospinal fluid, blood, urine) Main techniques direct agglutination: slides, cards latex agglutination: slides, cards immuno-chromatography: dipsticks
DR.T.V.RAO MD
12-10-2012
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. DIRECT METHODS - PROPAGATION REQUIRED Bacteriology and mycology
most agents can be propagated on culture media
Virology
most agents can be propagated on cells
Parasitology
monocellular organisms can be propagated in culture media
DR.T.V.RAO MD 12-10-2012
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PERSONNEL
The following directorial functions are :
1- interpretation , correlation , and communication of lab data 2- interaction with physicians and/or medical staff , patient , administration . 3- monitoring of standard of performance , QC , QI. 4- provision of education programs , planning , research. 5- ensuring sufficient personnel with adequate documented training and experience to meet the needs of the lab .
DR.T.V.RAO MD 12-10-2012
6- he/she must be decision-maker in the selection of all lab equipment's and supplies . 21
PROPAGATION: ADVANTAGES/DISADVANTAGES
Advantages
allows anti-microbial susceptibility testing
allows typing of the micro-organism allows storage of the strain
Disadvantages
depends upon the viability/condition of the agent takes time
DR.T.V.RAO MD 12-10-2012
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DETECTING ANTIBODIES
Precipitation
Agglutination Haemagglutination and haemagglutination inhibition Viral neutralization test Radio-immunoassays ELISA Immunofluorescence Immmunoblotting Immunochromatographic
DR.T.V.RAO MD 12-10-2012
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ANTIBODY DETECTION: ADVANTAGES/DISADVANTAGES
Advantages
inexpensive
easy to perform allows identification of
IgM (acute infection)
IgG (past infection) Disadvantages
delayed response (false negative results during sero-conversion window)
time of infection not always clear
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STANDARDS AND CHECKLISTS
Purposes:
Standards are the broad principles the laboratory must meet in order to achieve accreditation Checklists provide detailed requirements that inspectors use to determine whether laboratories meet the Standards
DR.T.V.RAO MD 12-10-2012
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LABORATORY INSPECTIONS: MAINTAINING BALANCE
Education Quality Improvement
DR.T.V.RAO MD
Regulatory Compliance
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THE STANDARDS FOR LABORATORY ACCREDITATION
Standard I Standard II Director / Head of the Divison Physical Facilities & Safety
Standard III
Standard IV
Quality Control and Performance Improvement
Inspection Requirements
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CHECKLISTS
Guide the inspection by assisting with the interpretation of desired Standards Provide guidelines for development of laboratory policies, procedures and processes Help ensure accurate, reliable test results Ensure a focus on patient safety
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Quality control and Quality improvement
The QC / QI program should be clearly defined and wellorganized. The QI program must provide the system design and evaluation of proper patient identification and preparation ; specimen collection ; preservation ; transportation ; storage before testing ; processing ; and accurate results reporting.
This system must ensure optimum patient specimen and integrity of the result throughout the pre-analytical , analytical , and post-analytical process.
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QI / QA
SUPERVISION
Judgment of the acceptability of QC data must be made at least monthly by the lab director . Because of many variables , the CAP makes no specific recommendations on the frequency of any additional assessment / review of QC data. There must be evidence of active review of records of instrument function , temp , and maintenance , for all routine procedures on all shifts.
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QI / QA
The lab must have documented system in operation to detect and correct significant clerical and analytical errors. One common method is review of results by a qualified person before release from the lab , but there is no requirement for supervisory review of all reported data. The selective use of delta checks also may be useful in detecting clerical errors in consecutive samples from the same patient. In computerized lab , there should be automatic alarm for improbable results.
DR.T.V.RAO MD 12-10-2012
SUPERVISION
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EFFECTS OF PRE-ANALYTICAL VARIABLES ON THE QUALITY OF LABORATORY TESTING
Modern laboratories around the world are now enjoying the benefit of decades of development in technology State of the Art" instrumentation are common in most laboratories Walkway, high throughput analyzers are employed for routine and specialized testing
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Programme Created by Dr.T.V.Rao MD for Medical Microbiologists
Email
[email protected]DR.T.V.RAO MD
12-10-2012
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